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A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2 | bioRxiv
<!DOCTYPE html> <html lang="en" dir="ltr" xmlns="http://www.w3.org/1999/xhtml" xmlns:mml="http://www.w3.org/1998/Math/MathML"> <head prefix="og: http://ogp.me/ns# article: http://ogp.me/ns/article# book: http://ogp.me/ns/book#" > <!--[if IE]><![endif]--> <link rel="dns-prefetch" href="//d33xdlntwy0kbs.cloudfront.net" /> <link rel="dns-prefetch" href="//www.google.com" /> <link rel="dns-prefetch" href="//scholar.google.com" /> <link rel="dns-prefetch" href="//www.googletagmanager.com" /> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <link rel="shortcut icon" href="https://www.biorxiv.org/sites/default/files/images/favicon.ico" type="image/vnd.microsoft.icon" /> <meta name="viewport" content="width=device-width, initial-scale=1" /> <link rel="alternate" type="application/pdf" title="Full Text (PDF)" href="/content/10.1101/2022.03.08.483529v1.full.pdf" /> <link rel="alternate" type="text/plain" title="Full Text (Plain)" href="/content/10.1101/2022.03.08.483529v1.full.txt" /> <meta name="type" content="article" /> <meta name="category" content="article" /> <meta name="HW.identifier" content="/biorxiv/early/2022/03/09/2022.03.08.483529.atom" /> <meta name="HW.pisa" content="biorxiv;2022.03.08.483529v1" /> <meta name="DC.Format" content="text/html" /> <meta name="DC.Language" content="en" /> <meta name="DC.Title" content="A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2" /> <meta name="DC.Identifier" content="10.1101/2022.03.08.483529" /> <meta name="DC.Date" content="2022-03-09" /> <meta name="DC.Publisher" content="Cold Spring Harbor Laboratory" /> <meta name="DC.Rights" content="© 2022, Posted by Cold Spring Harbor Laboratory. The copyright holder for this pre-print is the author. All rights reserved. The material may not be redistributed, re-used or adapted without the author's permission." /> <meta name="DC.AccessRights" content="restricted" /> <meta name="DC.Description" content="During development cell fates are specified by tightly controlled gene expression programs. PBX TALE transcription factors control gene regulatory networks (GRN) that direct vertebrate tissue patterning and organ morphogenesis. How PBX1/2 proteins acquire context-specific functions, despite widespread embryonic expression of Pbx1/2 , remains elusive. In mouse limb buds, mesenchymal-specific loss of PBX1/2 or of the transcriptional regulator HAND2 results in similar phenotypes, suggesting that PBX1/2- and HAND2-dependent programs converge to control limb development. To investigate this scenario, we combined tissue-specific and temporally-controlled mutagenesis with multi-omics approaches using the murine hindlimb bud as a model. We reconstructed a GRN collaboratively directed by PBX1/2 and HAND2, demonstrating that Pbx1 - Hand2 genetically interact in vivo during hindlimb patterning, with PBX1 concomitantly acting as an upstream regulator of Hand2 . At organismal-level resolution the GRN is active within restricted subsets of posterior-proximal hindlimb mesenchymal cells, wherein Pbx1/2 and Hand2 are co-expressed with their target genes. Genome-wide profiling of PBX1 binding across multiple tissues further revealed that HAND2 selects a subset of PBX-bound regions to impart limb patterning functionality. This research elucidates mechanisms underlying limb bud-specific functions by PBX1/2, while informing general principles by which promiscuous transcription factors cooperate with select cofactors to instruct distinct developmental programs. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="DC.Contributor" content="Marta Losa" /> <meta name="DC.Contributor" content="Iros Barozzi" /> <meta name="DC.Contributor" content="Marco Osterwalder" /> <meta name="DC.Contributor" content="Peyman Zarrineh" /> <meta name="DC.Contributor" content="Jean Denis Benazet" /> <meta name="DC.Contributor" content="Brandon Chacon" /> <meta name="DC.Contributor" content="Ausra Girdziusaite" /> <meta name="DC.Contributor" content="Angela Morabito" /> <meta name="DC.Contributor" content="Jianjian Zhu" /> <meta name="DC.Contributor" content="Susan Mackem" /> <meta name="DC.Contributor" content="Terence D. Capellini" /> <meta name="DC.Contributor" content="Nicoletta Bobola" /> <meta name="DC.Contributor" content="Diane Dickel" /> <meta name="DC.Contributor" content="Aimee Zuniga" /> <meta name="DC.Contributor" content="Axel Visel" /> <meta name="DC.Contributor" content="Rolf Zeller" /> <meta name="DC.Contributor" content="Licia Selleri" /> <meta name="article:published_time" content="2022-03-09" /> <meta name="article:section" content="New Results" /> <meta name="citation_title" content="A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2" /> <meta name="citation_abstract" lang="en" content="<h3>ABSTRACT</h3> <p>During development cell fates are specified by tightly controlled gene expression programs. PBX TALE transcription factors control gene regulatory networks (GRN) that direct vertebrate tissue patterning and organ morphogenesis. How PBX1/2 proteins acquire context-specific functions, despite widespread embryonic expression of <i>Pbx1/2</i>, remains elusive. In mouse limb buds, mesenchymal-specific loss of PBX1/2 or of the transcriptional regulator HAND2 results in similar phenotypes, suggesting that PBX1/2- and HAND2-dependent programs converge to control limb development. To investigate this scenario, we combined tissue-specific and temporally-controlled mutagenesis with multi-omics approaches using the murine hindlimb bud as a model. We reconstructed a GRN collaboratively directed by PBX1/2 and HAND2, demonstrating that <i>Pbx1</i>-<i>Hand2</i> genetically interact <i>in vivo</i> during hindlimb patterning, with PBX1 concomitantly acting as an upstream regulator of <i>Hand2</i>. At organismal-level resolution the GRN is active within restricted subsets of posterior-proximal hindlimb mesenchymal cells, wherein <i>Pbx1/2</i> and <i>Hand2</i> are co-expressed with their target genes. Genome-wide profiling of PBX1 binding across multiple tissues further revealed that HAND2 selects a subset of PBX-bound regions to impart limb patterning functionality. This research elucidates mechanisms underlying limb bud-specific functions by PBX1/2, while informing general principles by which promiscuous transcription factors cooperate with select cofactors to instruct distinct developmental programs.</p>" /> <meta name="citation_journal_title" content="bioRxiv" /> <meta name="citation_publisher" content="Cold Spring Harbor Laboratory" /> <meta name="citation_publication_date" content="2022/01/01" /> <meta name="citation_mjid" content="biorxiv;2022.03.08.483529v1" /> <meta name="citation_id" content="2022.03.08.483529v1" /> <meta name="citation_public_url" content="https://www.biorxiv.org/content/10.1101/2022.03.08.483529v1" /> <meta name="citation_abstract_html_url" content="https://www.biorxiv.org/content/10.1101/2022.03.08.483529v1.abstract" /> <meta name="citation_full_html_url" content="https://www.biorxiv.org/content/10.1101/2022.03.08.483529v1.full" /> <meta name="citation_pdf_url" content="https://www.biorxiv.org/content/biorxiv/early/2022/03/09/2022.03.08.483529.full.pdf" /> <meta name="citation_doi" content="10.1101/2022.03.08.483529" /> <meta name="citation_num_pages" content="42" /> <meta name="citation_article_type" content="Article" /> <meta name="citation_section" content="New Results" /> <meta name="citation_firstpage" content="2022.03.08.483529" /> <meta name="citation_author" content="Marta Losa" /> <meta name="citation_author_institution" content="Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco" /> <meta name="citation_author" content="Iros Barozzi" /> <meta name="citation_author_institution" content="Center for Cancer Research, Medical University of Vienna" /> <meta name="citation_author" content="Marco Osterwalder" /> <meta name="citation_author_institution" content="Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory" /> <meta name="citation_author_institution" content="Department for Biomedical Research (DBMR), University of Bern" /> <meta name="citation_author_institution" content="Department of Cardiology, Bern University Hospital" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-1969-2313" /> <meta name="citation_author" content="Peyman Zarrineh" /> <meta name="citation_author_institution" content="School of Medical Sciences, University of Manchester" /> <meta name="citation_author" content="Jean Denis Benazet" /> <meta name="citation_author_institution" content="Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco" /> <meta name="citation_author" content="Brandon Chacon" /> <meta name="citation_author_institution" content="Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco" /> <meta name="citation_author" content="Ausra Girdziusaite" /> <meta name="citation_author_institution" content="Developmental Genetics, Department Biomedicine, University of Basel" /> <meta name="citation_author" content="Angela Morabito" /> <meta name="citation_author_institution" content="Developmental Genetics, Department Biomedicine, University of Basel" /> <meta name="citation_author" content="Jianjian Zhu" /> <meta name="citation_author_institution" content="Cancer and Developmental Biology Laboratory, Center for Cancer Research, National Cancer Institute" /> <meta name="citation_author" content="Susan Mackem" /> <meta name="citation_author_institution" content="Cancer and Developmental Biology Laboratory, Center for Cancer Research, National Cancer Institute" /> <meta name="citation_author" content="Terence D. Capellini" /> <meta name="citation_author_institution" content="Department of Human Evolutionary Biology, Harvard University" /> <meta name="citation_author_institution" content="Broad Institute of Harvard and MIT" /> <meta name="citation_author" content="Nicoletta Bobola" /> <meta name="citation_author_institution" content="School of Medical Sciences, University of Manchester" /> <meta name="citation_author" content="Diane Dickel" /> <meta name="citation_author_institution" content="Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory" /> <meta name="citation_author" content="Aimee Zuniga" /> <meta name="citation_author_institution" content="Developmental Genetics, Department Biomedicine, University of Basel" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-9953-3637" /> <meta name="citation_author" content="Axel Visel" /> <meta name="citation_author_institution" content="Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory" /> <meta name="citation_author_institution" content="US Department of Energy Joint Genome Institute, Lawrence Berkeley National Laboratory" /> <meta name="citation_author_institution" content="School of Natural Sciences, University of California" /> <meta name="citation_author" content="Rolf Zeller" /> <meta name="citation_author_institution" content="Developmental Genetics, Department Biomedicine, University of Basel" /> <meta name="citation_author_orcid" content="http://orcid.org/0000-0002-3186-7403" /> <meta name="citation_author" content="Licia Selleri" /> <meta name="citation_author_institution" content="Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco" /> <meta name="citation_author_email" content="licia.selleri@ucsf.edu" /> <meta name="citation_reference" content="Amândio AR, Beccari L, Lopez-Delisle L, Mascrez B, Zakany J, Gitto S, Duboule D. 2021. Sequential in cis mutagenesis in vivo reveals various functions for CTCF sites at the mouse HoxD cluster. Genes and Development 35: 1490–1509. http://genesdev.cshlp.org/content/35/21-22/1490.full." /> <meta name="citation_reference" content="Amin S, Donaldson IJ, Zannino DA, Hensman J, Rattray M, Losa M, Spitz F, Ladam F, Sagerström C, Bobola N. 2015. Hoxa2 Selectively Enhances Meis Binding to Change a Branchial Arch Ground State. Developmental Cell 32: 265–277. http://www.cell.com/article/S1534580714008454/fulltext." /> <meta name="citation_reference" content="Bardet AF, He Q, Zeitlinger J, Stark A. 2011. A computational pipeline for comparative ChIP-seq analyses. Nature Protocols 2012 7:1 7: 45–61. https://www.nature.com/articles/nprot.2011.420." /> <meta name="citation_reference" content="Bensoussan-Trigano V, Lallemand Y, saint Cloment C, Robert B. 2011. Msx1 and Msx2 in limb mesenchyme modulate digit number and identity. Developmental Dynamics 240: 1190–1202. https://pubmed.ncbi.nlm.nih.gov/21465616/." /> <meta name="citation_reference" content="Berge D ten, Brouwer A, Korving J, Martin JF, Meijlink F. 1998. Prx1 and Prx2 in skeletogenesis: roles in the craniofacial region, inner ear and limbs. Development 125: 3831–3842." /> <meta name="citation_reference" content="Berkes CA, Bergstrom DA, Penn BH, Seaver KJ, Knoepfler PS, Tapscott SJ. 2004. Pbx marks genes for activation by MyoD indicating a role for a homeodomain protein in establishing myogenic potential. Molecular cell 14: 465–477. https://pubmed.ncbi.nlm.nih.gov/15149596/." /> <meta name="citation_reference" content="Bobola N, Merabet S. 2017. Homeodomain proteins in action: similar DNA binding preferences, highly variable connectivity. Current opinion in genetics & development 43: 1–8. https://pubmed.ncbi.nlm.nih.gov/27768937/" /> <meta name="citation_reference" content="Bolt CC, Duboule D. 2020. The regulatory landscapes of developmental genes. Development 147." /> <meta name="citation_reference" content="Bonev B, Cohen NM, Szabo Q, Fritsch L, Papadopoulos GL, Lubling Y, Xu X, Lv X, Hugnot J-P, Tanay A, et al. 2017. Multiscale 3D Genome Rewiring during Mouse Neural Development. Cell 171: 557. https://pmc/articles/PMC5651218/." /> <meta name="citation_reference" content="Brendolan A, Ferretti E, Salsi V, Moses K, Quaggin S, Blasi F, Cleary ML, Selleri L. 2005. A Pbx1-dependent genetic and transcriptional network regulates spleen ontogeny. Development 132: 3113–3126. https://pubmed.ncbi.nlm.nih.gov/15944191/." /> <meta name="citation_reference" content="Bridoux L, Zarrineh P, Mallen J, Phuycharoen M, Latorre V, Ladam F, Losa M, Baker SM, Sagerstrom C, Mace KA, et al. 2020. HOX paralogs selectively convert binding of ubiquitous transcription factors into tissue-specific patterns of enhancer activation. PLOS Genetics 16: e1009162. https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1009162 (Accessed July 19, 2021)." /> <meta name="citation_reference" content="Buenrostro JD, Giresi PG, Zaba LC, Chang HY, Greenleaf WJ. 2013. Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nature Methods 2013 10:12 10: 1213–1218. https://www.nature.com/articles/nmeth.2688." /> <meta name="citation_reference" content="Bürglin TR, Affolter M. 2015. Homeodomain proteins: an update. Chromosoma 2015 125:3 125: 497–521. https://link.springer.com/article/10.1007/s00412-015-0543-8." /> <meta name="citation_reference" content="Cao J, Spielmann M, Qiu X, Huang X, Ibrahim DM, Hill AJ, Zhang F, Mundlos S, Christiansen L, Steemers FJ, et al. 2019. The single-cell transcriptional landscape of mammalian organogenesis. Nature 2019 566:7745 566: 496–502." /> <meta name="citation_reference" content="Capellini T, Vaccari G, Ferretti E, Fantini S, He M, Pellegrini M, Quintana L, di Giacomo G, Sharpe J, Selleri L, et al. 2010. Scapula development is governed by genetic interactions of Pbx1 with its family members and with Emx2 via their cooperative control of Alx1. Development 137: 2559–2569. https://pubmed.ncbi.nlm.nih.gov/20627960/." /> <meta name="citation_reference" content="Capellini TD, di Giacomo G, Salsi V, Brendolan A, Ferretti E, Srivastava D, Zappavigna V, Selleri L. 2006. Pbx1/Pbx2 requirement for distal limb patterning is mediated by the hierarchical control of Hox gene spatial distribution and Shh expression. Development 133: 2263–2273. http://www.ncbi.nlm.nih.gov/pubmed/16672333." /> <meta name="citation_reference" content="Capellini TD, Handschuh K, Quintana L, Ferretti E, Giacomo G di, Fantini S, Vaccari G, Clarke SL, Wenger AM, Bejerano G, et al. 2011a. Control of pelvic girdle development by genes of the Pbx family and Emx2. Developmental Dynamics 240: 1173–1189. https://pubmed.ncbi.nlm.nih.gov/21455939/." /> <meta name="citation_reference" content="Capellini TD, Zappavigna V, Selleri L. 2011b. Pbx homeodomain proteins: TALEnted regulators of limb patterning and outgrowth. Developmental Dynamics 240: 1063–1086. http://www.ncbi.nlm.nih.gov/pubmed/21416555." /> <meta name="citation_reference" content="Capellini TD, Zewdu R, di Giacomo G, Asciutti S, Kugler JE, di Gregorio A, Selleri L. 2008. Pbx1/Pbx2 govern axial skeletal development by controlling Polycomb and Hox in mesoderm and Pax1/Pax9 in sclerotome. Developmental Biology 321: 500–514. http://www.ncbi.nlm.nih.gov/pubmed/18691704." /> <meta name="citation_reference" content="Chan KKK, Zhang J, Chia NY, Chan YS, Sim HS, Tan KS, Oh SKW, Ng HH, Choo ABH. 2009. KLF4 and PBX1 directly regulate NANOG expression in human embryonic stem cells. Stem cells 27: 2114–2125. https://pubmed.ncbi.nlm.nih.gov/19522013/." /> <meta name="citation_reference" content="Charite J, McFadden DG, Olson EN. 2000. The bHLH transcription factor dHAND controls Sonic hedgehog expression and establishment of the zone of polarizing activity during limb development. Development 127: 2461–2470." /> <meta name="citation_reference" content="Chen Y, Gridley T. 2013. Compensatory regulation of the Snai1 and Snai2 genes during chondrogenesis. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research 28: 1412. https://pmc/articles/PMC3663919/." /> <meta name="citation_reference" content="Chiang C, Litingtung Y, Harris MP, Simandl BK, Li Y, Beachy PA, Fallon JF. 2001. Manifestation of the Limb Prepattern: Limb Development in the Absence of Sonic Hedgehog Function. Developmental Biology 236: 421–435." /> <meta name="citation_reference" content="Choe SK, Ladam F, Sagerström CG. 2014. TALE factors poise promoters for activation by Hox proteins. Developmental cell 28: 203–211. https://pubmed.ncbi.nlm.nih.gov/24480644/." /> <meta name="citation_reference" content="Choe SK, Lu P, Nakamura M, Lee J, Sagerström CG. 2009. Meis cofactors control HDAC and CBP accessibility at Hox-regulated promoters during zebrafish embryogenesis. Developmental cell 17: 561–567. https://pubmed.ncbi.nlm.nih.gov/19853569/." /> <meta name="citation_reference" content="Creuzet S, Couly G, le Douarin NM. 2005. Patterning the neural crest derivatives during development of the vertebrate head: insights from avian studies. Journal of anatomy 207: 447–459. https://pubmed.ncbi.nlm.nih.gov/16313387/." /> <meta name="citation_reference" content="Darbellay F, Duboule D. 2016. Topological Domains, Metagenes, and the Emergence of Pleiotropic Regulations at Hox Loci. Current topics in developmental biology 116: 299–314." /> <meta name="citation_reference" content="de Frutos CA, Vega S, Manzanares M, Flores JM, Huertas H, Martínez-Frías ML, Nieto MA. 2007. Snail1 is a transcriptional effector of FGFR3 signaling during chondrogenesis and achondroplasias. Developmental cell 13: 872–883. https://pubmed.ncbi.nlm.nih.gov/18061568/." /> <meta name="citation_reference" content="de Kumar B, Parker HJ, Paulson A, Parrish ME, Pushel I, Singh NP, Zhang Y, Slaughter BD, Unruh JR, Florens L, et al. 2017. HOXA1 and TALE proteins display cross-regulatory interactions and form a combinatorial binding code on HOXA1 targets. Genome research 27: 1501–1512. https://pubmed.ncbi.nlm.nih.gov/28784834/." /> <meta name="citation_reference" content="Delgado I, Giovinazzo G, Temiño S, Gauthier Y, Balsalobre A, Drouin J, Torres M. 2021. Control of mouse limb initiation and antero-posterior patterning by Meis transcription factors. Nature Communications 2021 12:1 12: 1–13. https://www.nature.com/articles/s41467-021-23373-9." /> <meta name="citation_reference" content="Delgado I, López-Delgado AC, Roselló-Díez A, Giovinazzo G, Cadenas V, Fernández-de-Manuel L, Sánchez-Cabo F, Anderson MJ, Lewandoski M, Torres M. 2020. Proximo-distal positional information encoded by an Fgf-regulated gradient of homeodomain transcription factors in the vertebrate limb. Science Advances 6: eaaz0742. https://advances.sciencemag.org/content/6/23/eaaz0742." /> <meta name="citation_reference" content="DeMare LE, Leng J, Cotney J, Reilly SK, Yin J, Sarro R, Noonan JP. 2013. The genomic landscape of cohesin-associated chromatin interactions. Genome research 23: 1224–1234. http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=3730097&tool=pmcentrez&rendertype=abstract." /> <meta name="citation_reference" content="Desanlis I, Paul R, Kmita M. 2020. Transcriptional Trajectories in Mouse Limb Buds Reveal the Transition from Anterior-Posterior to Proximal-Distal Patterning at Early Limb Bud Stage. Journal of Developmental Biology 2020, Vol 8, Page 31 8: 31." /> <meta name="citation_reference" content="Deschamps J, Duboule D. 2017. Embryonic timing, axial stem cells, chromatin dynamics, and the Hox clock. Genes & development 31: 1406–1416. https://pubmed.ncbi.nlm.nih.gov/28860158/." /> <meta name="citation_reference" content="di Giacomo G, Koss M, Capellini T, Brendolan A, Pöpperl H, Selleri L. 2006. Spatio-temporal expression of Pbx3 during mouse organogenesis. Gene expression patterns : GEP 6: 747–757. https://pubmed.ncbi.nlm.nih.gov/16434237/." /> <meta name="citation_reference" content="Dixon JR, Selvaraj S, Yue F, Kim A, Li Y, Shen Y, Hu M, Liu JS, Ren B. 2012. Topological Domains in Mammalian Genomes Identified by Analysis of Chromatin Interactions. Nature 485: 376." /> <meta name="citation_reference" content="Donaghey J, Thakurela S, Charlton J, Chen JS, Smith ZD, Gu H, Pop R, Clement K, Stamenova EK, Karnik R, et al. 2018. Genetic determinants and epigenetic effects of pioneer-factor occupancy. Nature Genetics 2017 50:2 50: 250–258. https://www.nature.com/articles/s41588-017-0034-3." /> <meta name="citation_reference" content="Donovan BT, Huynh A, Ball DA, Patel HP, Poirier MG, Larson DR, Ferguson ML, Lenstra TL. 2019. Live-cell imaging reveals the interplay between transcription factors, nucleosomes, and bursting. The EMBO Journal 38: e100809. https://onlinelibrary.wiley.com/doi/full/10.15252/embj.2018100809." /> <meta name="citation_reference" content="Fabre PJ, Leleu M, Mascrez B, lo Giudice Q, Cobb J, Duboule D. 2018. Heterogeneous combinatorial expression of Hoxd genes in single cells during limb development. BMC Biology 16: 1–15. https://bmcbiol.biomedcentral.com/articles/10.1186/s12915-018-0570-z." /> <meta name="citation_reference" content="Fernández-Terán MA, Hinchliffe JR, Ros MA. 2006. Birth and death of cells in limb development: A mapping study. Developmental Dynamics 235: 2521–2537." /> <meta name="citation_reference" content="Ferretti E, Li B, Zewdu R, Wells V, Hebert JM, Karner C, Anderson MJ, Williams T, Dixon J, Dixon MJ, et al. 2011. A Conserved Pbx-Wnt-p63-Irf6 Regulatory Module Controls Face Morphogenesis by Promoting Epithelial Apoptosis. Developmental Cell 21: 627–641." /> <meta name="citation_reference" content="citation_title=PLoS Genetics;citation_author=A. Galli;citation_author=D. Robay;citation_author=M. Osterwalder;citation_author=X. Bao;citation_author=J-D. Bénazet;citation_author=M. Tariq;citation_author=R. Paro;citation_author=S. Mackem;citation_author=R. Zeller;citation_pages=e1000901;citation_volume=6;citation_year=2010" /> <meta name="citation_reference" content="Garcia-Alonso L, Holland CH, Ibrahim MM, Turei D, Saez-Rodriguez J. 2019. Benchmark and integration of resources for the estimation of human transcription factor activities. Genome research 29: 1363–1375. https://pubmed.ncbi.nlm.nih.gov/31340985/." /> <meta name="citation_reference" content="Golonzhka O, Nord A, Tang PLF, Lindtner S, Ypsilanti AR, Ferretti E, Visel A, Selleri L, Rubenstein JLR. 2015. Pbx Regulates Patterning of the Cerebral Cortex in Progenitors and Postmitotic Neurons. Neuron 88: 1192–1207." /> <meta name="citation_reference" content="Grebbin BM, Hau AC, Groß A, Anders-Maurer M, Schramm J, Koss M, Wille C, Mittelbronn M, Selleri L, Schulte D. 2016. PBX1 is required for adult subventricular zone neurogenesis. Development (Cambridge) 143: 2281–2291." /> <meta name="citation_reference" content="Grebbin BM, Schulte D. 2017. PBX1 as Pioneer Factor: A Case Still Open. Frontiers in Cell and Developmental Biology 5." /> <meta name="citation_reference" content="Heinz S, Benner C, Spann N, Bertolino E, Lin Y, Laslo P, Cheng J, Murre C, Singh H, Glass C. 2010. Simple combinations of lineage-determining transcription factors prime cis-regulatory elements required for macrophage and B cell identities. Molecular cell 38: 576–589. https://pubmed.ncbi.nlm.nih.gov/20513432/." /> <meta name="citation_reference" content="Hurtado R, Zewdu R, Mtui J, Liang C, Aho R, Kurylo C, Selleri L, Herzlinger D. 2015. Pbx1-dependent control of VMC differentiation kinetics underlies gross renal vascular patterning. Development (Cambridge) 142: 2653–2664." /> <meta name="citation_reference" content="Iwafuchi-Doi M, Zaret KS. 2014. Pioneer transcription factors in cell reprogramming. Genes & development 28: 2679–2692. https://pubmed.ncbi.nlm.nih.gov/25512556/." /> <meta name="citation_reference" content="Jürgens AS, Kolanczyk M, Moebest DCC, Zemojtel T, Lichtenauer U, Duchniewicz M, Gantert MP, Hecht J, Hattenhorst U, Burdach S, et al. 2009. PBX1 is dispensable for neural commitment of RA-treated murine ES cells. In vitro cellular & developmental biology Animal 45: 252–263. https://pubmed.ncbi.nlm.nih.gov/19148706/." /> <meta name="citation_reference" content="Kelly NH, Huynh NPT, Guilak F. 2020. Single cell RNA-sequencing reveals cellular heterogeneity and trajectories of lineage specification during murine embryonic limb development. Matrix Biology 89: 1–10." /> <meta name="citation_reference" content="Kmita M, Tarchini B, Zàkàny J, Logan M, Tabin CJ, Duboule D. 2005. Early developmental arrest of mammalian limbs lacking HoxA/HoxD gene function. Nature 2005 435:7045 435: 1113–1116. https://www.nature.com/articles/nature03648." /> <meta name="citation_reference" content="Koss M, Bolze A, Brendolan A, Saggese M, Capellini TD, Bojilova E, Boisson B, Prall OWJ, Elliott DA, Solloway M, et al. 2012. Congenital Asplenia in Mice and Humans with Mutations in a Pbx/Nkx2-5/p15 Module. Developmental Cell 22: 913–926." /> <meta name="citation_reference" content="Kothary R, Clapoff S, Darling S, Perry MD, Moran LA, Rossant J. 1989. Inducible expression of an hsp68-lacZ hybrid gene in transgenic mice. Development 105: 707–714. https://pubmed.ncbi.nlm.nih.gov/2557196/." /> <meta name="citation_reference" content="Kozhemyakina E, Ionescu A, Lassar AB. 2014. GATA6 Is a Crucial Regulator of Shh in the Limb Bud. PLOS Genetics 10: e1004072. https://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1004072." /> <meta name="citation_reference" content="Kuijper S, Beverdam A, Kroon C, Brouwer A, Candille S, Barsh G, Meijlink F. 2005. Genetics of shoulder girdle formation: roles of Tbx15 and aristaless-like genes. Development 132: 1601–1610." /> <meta name="citation_reference" content="Kvon EZ, Zhu Y, Kelman G, Visel A, Dickel DE, Pennacchio LA. 2020. Comprehensive In Vivo Interrogation Reveals Phenotypic Impact of Human Enhancer Variants. Cell 180. https://doi.org/10.1016/j.cell.2020.02.031." /> <meta name="citation_reference" content="Lee TI, Young RA. 2013. Transcriptional Regulation and its Misregulation in Disease. Cell 152: 1237. https://pmc/articles/PMC3640494/." /> <meta name="citation_reference" content="Lettice LA, Williamson I, Devenney PS, Kilanowski F, Dorin J, Hill RE. 2014. Development of five digits is controlled by a bipartite long-range cis-regulator. Development 141: 1715–1725." /> <meta name="citation_reference" content="Lettice LA, Williamson I, Wiltshire JH, Peluso S, Devenney PS, Hill AE, Essafi A, Hagman J, Mort R, Grimes G, et al. 2012. Opposing Functions of the ETS Factor Family Define Shh Spatial Expression in Limb Buds and Underlie Polydactyly. Developmental Cell 22: 459. https://pmc/articles/PMC3314984/." /> <meta name="citation_reference" content="Lickwar CR, Mueller F, Hanlon SE, McNally JG, Lieb JD. 2012. Genome-wide protein–DNA binding dynamics suggest a molecular clutch for transcription factor function. Nature 2012 484:7393 484: 251–255. https://www.nature.com/articles/nature10985." /> <meta name="citation_reference" content="Liu Y, Ma L, Wu L, Luo W, Kundu R, Sangiorgi F, Snead M, Maxson R. 1994. Regulation of the Msx2 homeobox gene during mouse embryogenesis: a transgene with 439 bp of 5’ flanking sequence is expressed exclusively in the apical ectodermal ridge of the developing limb. Mechanisms of development 48: 187–197. https://pubmed.ncbi.nlm.nih.gov/7893602/." /> <meta name="citation_reference" content="Losa M, Latorre V, Andrabi M, Ladam F, Sagerström C, Novoa A, Zarrineh P, Bridoux L, Hanley NA, Mallo M, et al. 2017. A tissue-specific, Gata6-driven transcriptional program instructs remodeling of the mature arterial tree. eLife 6." /> <meta name="citation_reference" content="Losa M, Risolino M, Li B, Hart J, Quintana L, Grishina I, Yang H, Choi IF, Lewicki P, Khan S, et al. 2018. Face morphogenesis is promoted by Pbx-dependent EMT via regulation of Snail1 during frontonasal prominence fusion. Development 145." /> <meta name="citation_reference" content="Lowe LA, Yamada S, Kuehn MR. 2000. HoxB6-Cre Transgenic Mice Express Cre Recombinase in Extra-Embryonic Mesoderm, in Lateral Plate and Limb Mesoderm and at the Midbrain/Hindbrain Junction. Genesis 118–120." /> <meta name="citation_reference" content="Malkmus J, Ramos Martins L, Jhanwar S, Kircher B, Palacio V, Sheth R, Leal F, Duchesne A, Lopez-Rios J, Peterson KA, et al. 2021. Spatial regulation by multiple Gremlin1 enhancers provides digit development with cis-regulatory robustness and evolutionary plasticity. Nature communications 12. https://pubmed.ncbi.nlm.nih.gov/34548488/." /> <meta name="citation_reference" content="Mann RS, Lelli KM, Joshi R. 2009. Hox specificity unique roles for cofactors and collaborators. Current topics in developmental biology 88: 63–101. https://pubmed.ncbi.nlm.nih.gov/19651302/." /> <meta name="citation_reference" content="Mao J, McGlinn E, Huang P, Tabin CJ, McMahon AP. 2009. Fgf-Dependent Etv4/5 Activity Is Required for Posterior Restriction of Sonic hedgehog and Promoting Outgrowth of the Vertebrate Limb. Developmental Cell 16: 600–606." /> <meta name="citation_reference" content="Mariani L, Guo X, Menezes NA, Drozd AM, Çakal SD, Wang Q, Ferretti E. 2021. A TALE/HOX code unlocks WNT signalling response towards paraxial mesoderm. Nature communications 12. https://pubmed.ncbi.nlm.nih.gov/34446717/." /> <meta name="citation_reference" content="McCulley DJ, Wienhold MD, Hines EA, Hacker TA, Rogers A, Pewowaruk RJ, Zewdu R, Chesler NC, Selleri L, Sun X. 2018. PBX transcription factors drive pulmonary vascular adaptation to birth. Journal of Clinical Investigation 128: 655–667." /> <meta name="citation_reference" content="McLean C, Bristor D, Hiller M, Clarke S, Schaar B, Lowe C, Wenger A, Bejerano G. 2010. GREAT improves functional interpretation of cis-regulatory regions. Nature biotechnology 28: 495–501. https://pubmed.ncbi.nlm.nih.gov/20436461/." /> <meta name="citation_reference" content="Mirny LA. 2010. Nucleosome-mediated cooperativity between transcription factors. Proceedings of the National Academy of Sciences of the United States of America 107: 22534–22539. https://www.pnas.org/content/107/52/22534." /> <meta name="citation_reference" content="Moens CB, Selleri L. 2006. Hox cofactors in vertebrate development. Developmental Biology." /> <meta name="citation_reference" content="Monti R, Barozzi I, Osterwalder M, Lee E, Kato M, Garvin TH, Plajzer-Frick I, Pickle CS, Akiyama JA, Afzal V, et al. 2017. Limb-Enhancer Genie: An accessible resource of accurate enhancer predictions in the developing limb. PLoS Computational Biology 13: e1005720. https://pmc/articles/PMC5578682/." /> <meta name="citation_reference" content="Morgunova E, Taipale J. 2017. Structural perspective of cooperative transcription factor binding. Current Opinion in Structural Biology 47: 1–8." /> <meta name="citation_reference" content="Moyle-Heyrman G, Tims HS, Widom J. 2011. Structural constraints in collaborative competition of transcription factors against the nucleosome. Journal of molecular biology 412: 634–646. https://pubmed.ncbi.nlm.nih.gov/21821044/." /> <meta name="citation_reference" content="Nakamura E, Hata K, Takahata Y, Kurosaka H, Abe M, Abe T, Kihara M, Komori T, Kobayashi S, Murakami T, et al. 2021. Zfhx4 regulates endochondral ossification as the transcriptional platform of Osterix in mice. Communications Biology 2021 4:1 4: 1–11. https://www.nature.com/articles/s42003-021-02793-9." /> <meta name="citation_reference" content="Nguyen M, Zhu J, Nakamura E, Bao X, Mackem S. 2009. Tamoxifen-dependent, inducible Hoxb6CreERT recombinase function in lateral plate and limb mesoderm, CNS isthmic organizer, posterior trunk neural crest, hindgut, and tailbud. Developmental dynamics : an official publication of the American Association of Anatomists 238: 467–474. https://pubmed.ncbi.nlm.nih.gov/19161221/." /> <meta name="citation_reference" content="Osterwalder M, Speziale D, Shoukry M, Mohan R, Ivanek R, Kohler M, Beisel C, Wen X, Scales SJ, Christoffels VM, et al. 2014. HAND2 targets define a network of transcriptional regulators that compartmentalize the early limb bud mesenchyme. Developmental cell 31: 345–357. http://www.ncbi.nlm.nih.gov/pubmed/25453830." /> <meta name="citation_reference" content="Osterwalder M, Tran S, Hunter RD, Meky EM, von Maydell K, Harrington AN, Godoy J, Novak CS, Plajzer-Frick I, Zhu Y, et al. 2022. Characterization of Mammalian In Vivo Enhancers Using Mouse Transgenesis and CRISPR Genome Editing. In Methods in Molecular Biology, Vol. 2403 of, pp. 147–186, Humana, New York, NY https://link.springer.com/protocol/10.1007/978-1-0716-1847-9_11." /> <meta name="citation_reference" content="Parker HJ, de Kumar B, Green SA, Prummel KD, Hess C, Kaufman CK, Mosimann C, Wiedemann LM, Bronner ME, Krumlauf R. 2019. A Hox-TALE regulatory circuit for neural crest patterning is conserved across vertebrates. Nature communications 10. https://pubmed.ncbi.nlm.nih.gov/30867425/." /> <meta name="citation_reference" content="Penkov D, SanMartín DM, Fernandez-Díaz LC, Rosselló CA, Torroja C, Sánchez-Cabo F, Warnatz HJ, Sultan M, Yaspo ML, Gabrieli A, et al. 2013. Analysis of the DNA-Binding Profile and Function of TALE Homeoproteins Reveals Their Specialization and Specific Interactions with Hox Genes/Proteins. Cell Reports 3: 1321–1333." /> <meta name="citation_reference" content="Pöpperl H, Bienz M, Studer M, Chan SK, Aparicio S, Brenner S, Mann RS, Krumlauf R. 1995. Segmental expression of Hoxb-1 is controlled by a highly conserved autoregulatory loop dependent upon exd/pbx. Cell 81: 1031–1042. https://pubmed.ncbi.nlm.nih.gov/7600572/." /> <meta name="citation_reference" content="Rhee JW, Arata A, Selleri L, Jacobs Y, Arata S, Onimaru H, Cleary ML. 2004. Pbx3 Deficiency Results in Central Hypoventilation. The American Journal of Pathology 165: 1343. https://pmc/articles/PMC1618620/." /> <meta name="citation_reference" content="Royle S, Tabin C, Young J. 2021. Limb positioning and initiation: An evolutionary context of pattern and formation. Developmental dynamics : an official publication of the American Association of Anatomists." /> <meta name="citation_reference" content="Ryoo HD, Mann RS. 1999. The control of trunk Hox specificity and activity by Extradenticle. Genes & development 13: 1704–1716. https://pubmed.ncbi.nlm.nih.gov/10398683/." /> <meta name="citation_reference" content="Sagai T, Hosoya M, Mizushina Y, Tamura M, Shiroishi T. 2005. Elimination of a long-range cis-regulatory module causes complete loss of limb-specific Shh expression and truncation of the mouse limb. Development 132: 797–803. http://www-gsd.lbl.gov/vista/index.shtml." /> <meta name="citation_reference" content="Sagerström CG. 2004. PbX marks the spot. Developmental cell 6: 737–738. https://pubmed.ncbi.nlm.nih.gov/15177017/." /> <meta name="citation_reference" content="Selleri L, Depew MJ, Jacobs Y, Chanda K, Tsang KY, Cheah KSE, Rubenstein JLR, O’Gorman S, Cleary ML. 2001. Requirement for Pbx1 in skeletal patterning and programming chondrocyte proliferation and differentiation. Development 128: 3543–3557." /> <meta name="citation_reference" content="Selleri L, DiMartino J, van Deursen J, Brendolan A, Sanyal M, Boon E, Capellini T, Smith KS, Rhee J, Pöpperl H, et al. 2004. The TALE Homeodomain Protein Pbx2 Is Not Essential for Development and Long-Term Survival. Molecular and Cellular Biology 24: 5324–5331. https://pubmed.ncbi.nlm.nih.gov/15169896/." /> <meta name="citation_reference" content="Selleri L, Zappavigna V, Ferretti E. 2019. ‘Building a perfect body’: Control of vertebrate organogenesis by PBX-dependent regulatory networks. Genes and Development 33: 258–275. https://pubmed.ncbi.nlm.nih.gov/30824532/." /> <meta name="citation_reference" content="Sonnet W, Rezsöhazy R, Donnay I. 2012. Characterization of TALE genes expression during the first lineage segregation in mammalian embryos. Developmental dynamics : an official publication of the American Association of Anatomists 241: 1827–1839. https://pubmed.ncbi.nlm.nih.gov/22987645/." /> <meta name="citation_reference" content="Stankunas K, Shang C, Twu KY, Kao S-C, a Jenkins N, Copeland NG, Sanyal M, Selleri L, Cleary ML, Chang C-P. 2008. Pbx/Meis deficiencies demonstrate multigenetic origins of congenital heart disease. Circulation research 103: 702–709." /> <meta name="citation_reference" content="Sun X, Lewandoski M, Meyers EN, Liu Y-H, Maxson RE, Martin GR. 2000. Conditional inactivation of Fgf4 reveals complexity of signalling during limb bud development. Nature Genetics 2000 25:1 25: 83–86. https://www-nature-com.ucsf.idm.oclc.org/articles/ng0500_83." /> <meta name="citation_reference" content="te Welscher P, Fernandez-Teran M, Ros MA, Zeller R. 2002. Mutual genetic antagonism involving GLI3 and dHAND prepatterns the vertebrate limb bud mesenchyme prior to SHH signaling. Genes & Development 16: 421–426. http://genesdev.cshlp.org/content/16/4/421.full." /> <meta name="citation_reference" content="Tzchori I, Day TF, Carolan PJ, Zhao Y, Wassif CA, Li LQ, Lewandoski M, Gorivodsky M, Love PE, Porter FD, et al. 2009. LIM homeobox transcription factors integrate signaling events that control three-dimensional limb patterning and growth. Development 136: 1375–1385." /> <meta name="citation_reference" content="Wagner K, Mincheva A, Korn B, Lichter P, Pöpperl H. 2001. Pbx4, a new Pbx family member on mouse chromosome 8, is expressed during spermatogenesis. Mechanisms of Development 103: 127–131." /> <meta name="citation_reference" content="Welsh IC, Hart J, Brown JM, Hansen K, Marques MR, Aho RJ, Grishina I, Hurtado R, Herzlinger D, Ferretti E, et al. 2018. Pbx loss in cranial neural crest, unlike in epithelium, results in cleft palate only and a broader midface. Journal of Anatomy." /> <meta name="citation_reference" content="Xu B, Wellik DM. 2011. Axial Hox9 activity establishes the posterior field in the developing forelimb. Proceedings of the National Academy of Sciences of the United States of America 108: 4888–4891. https://pubmed.ncbi.nlm.nih.gov/21383175/." /> <meta name="citation_reference" content="Yin W, Mendoza L, Monzon-Sandoval J, Urrutia AO, Gutierrez H. 2021. Emergence of co-expression in gene regulatory networks. PLOS ONE 16: e0247671. https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0247671." /> <meta name="citation_reference" content="Zuniga A, Zeller R. 2020. Dynamic and self-regulatory interactions among gene regulatory networks control vertebrate limb bud morphogenesis. Current topics in developmental biology 139: 61–88. https://pubmed.ncbi.nlm.nih.gov/32450969/." /> <meta name="twitter:title" content="A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2" /> <meta name="twitter:site" content="@biorxivpreprint" /> <meta name="twitter:card" content="summary" /> <meta name="twitter:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="twitter:description" content="During development cell fates are specified by tightly controlled gene expression programs. PBX TALE transcription factors control gene regulatory networks (GRN) that direct vertebrate tissue patterning and organ morphogenesis. How PBX1/2 proteins acquire context-specific functions, despite widespread embryonic expression of Pbx1/2 , remains elusive. In mouse limb buds, mesenchymal-specific loss of PBX1/2 or of the transcriptional regulator HAND2 results in similar phenotypes, suggesting that PBX1/2- and HAND2-dependent programs converge to control limb development. To investigate this scenario, we combined tissue-specific and temporally-controlled mutagenesis with multi-omics approaches using the murine hindlimb bud as a model. We reconstructed a GRN collaboratively directed by PBX1/2 and HAND2, demonstrating that Pbx1 - Hand2 genetically interact in vivo during hindlimb patterning, with PBX1 concomitantly acting as an upstream regulator of Hand2 . At organismal-level resolution the GRN is active within restricted subsets of posterior-proximal hindlimb mesenchymal cells, wherein Pbx1/2 and Hand2 are co-expressed with their target genes. Genome-wide profiling of PBX1 binding across multiple tissues further revealed that HAND2 selects a subset of PBX-bound regions to impart limb patterning functionality. This research elucidates mechanisms underlying limb bud-specific functions by PBX1/2, while informing general principles by which promiscuous transcription factors cooperate with select cofactors to instruct distinct developmental programs. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-title" property="og:title" content="A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2" /> <meta name="og-url" property="og:url" content="https://www.biorxiv.org/content/10.1101/2022.03.08.483529v1" /> <meta name="og-site-name" property="og:site_name" content="bioRxiv" /> <meta name="og-description" property="og:description" content="During development cell fates are specified by tightly controlled gene expression programs. PBX TALE transcription factors control gene regulatory networks (GRN) that direct vertebrate tissue patterning and organ morphogenesis. How PBX1/2 proteins acquire context-specific functions, despite widespread embryonic expression of Pbx1/2 , remains elusive. In mouse limb buds, mesenchymal-specific loss of PBX1/2 or of the transcriptional regulator HAND2 results in similar phenotypes, suggesting that PBX1/2- and HAND2-dependent programs converge to control limb development. To investigate this scenario, we combined tissue-specific and temporally-controlled mutagenesis with multi-omics approaches using the murine hindlimb bud as a model. We reconstructed a GRN collaboratively directed by PBX1/2 and HAND2, demonstrating that Pbx1 - Hand2 genetically interact in vivo during hindlimb patterning, with PBX1 concomitantly acting as an upstream regulator of Hand2 . At organismal-level resolution the GRN is active within restricted subsets of posterior-proximal hindlimb mesenchymal cells, wherein Pbx1/2 and Hand2 are co-expressed with their target genes. Genome-wide profiling of PBX1 binding across multiple tissues further revealed that HAND2 selects a subset of PBX-bound regions to impart limb patterning functionality. This research elucidates mechanisms underlying limb bud-specific functions by PBX1/2, while informing general principles by which promiscuous transcription factors cooperate with select cofactors to instruct distinct developmental programs. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-type" property="og:type" content="article" /> <meta name="og-image" property="og:image" content="https://www.biorxiv.org/sites/default/files/images/biorxiv_logo_homepage7-5-small.png" /> <meta name="citation_date" content="2022-03-09" /> <link rel="alternate" type="application/vnd.ms-powerpoint" title="Powerpoint" href="/content/10.1101/2022.03.08.483529v1.ppt" /> <meta name="description" content="bioRxiv - the preprint server for biology, operated by Cold Spring Harbor Laboratory, a research and educational institution" /> <meta name="generator" content="Drupal 7 (http://drupal.org)" /> <link rel="canonical" 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data-apath="/biorxiv/early/2022/03/09/2022.03.08.483529.atom" data-hw-author-tooltip-instance="highwire_author_tooltip"><div class="highwire-cite highwire-cite-highwire-article highwire-citation-biorxiv-article-top clearfix has-author-tooltip" > <span class="biorxiv-article-type"> New Results </span> <h1 class="highwire-cite-title" id="page-title">A spatio-temporally constrained gene regulatory network directed by PBX1/2 acquires limb patterning specificity via HAND2</h1> <div class="highwire-cite-authors" ><span class="highwire-citation-authors"><span class="highwire-citation-author first" data-delta="0"><span class="nlm-given-names">Marta</span> <span class="nlm-surname">Losa</span></span>, <span class="highwire-citation-author" data-delta="1"><span class="nlm-given-names">Iros</span> <span class="nlm-surname">Barozzi</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="2"><a href="http://orcid.org/0000-0002-1969-2313" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Marco</span> <span class="nlm-surname">Osterwalder</span></span>, <span class="highwire-citation-author" data-delta="3"><span class="nlm-given-names">Peyman</span> <span class="nlm-surname">Zarrineh</span></span>, <span class="highwire-citation-author" data-delta="4"><span class="nlm-given-names">Jean Denis</span> <span class="nlm-surname">Benazet</span></span>, <span class="highwire-citation-author" data-delta="5"><span class="nlm-given-names">Brandon</span> <span class="nlm-surname">Chacon</span></span>, <span class="highwire-citation-author" data-delta="6"><span class="nlm-given-names">Ausra</span> <span class="nlm-surname">Girdziusaite</span></span>, <span class="highwire-citation-author" data-delta="7"><span class="nlm-given-names">Angela</span> <span class="nlm-surname">Morabito</span></span>, <span class="highwire-citation-author" data-delta="8"><span class="nlm-given-names">Jianjian</span> <span class="nlm-surname">Zhu</span></span>, <span class="highwire-citation-author" data-delta="9"><span class="nlm-given-names">Susan</span> <span class="nlm-surname">Mackem</span></span>, <span class="highwire-citation-author" data-delta="10"><span class="nlm-given-names">Terence D.</span> <span class="nlm-surname">Capellini</span></span>, <span class="highwire-citation-author" data-delta="11"><span class="nlm-given-names">Nicoletta</span> <span class="nlm-surname">Bobola</span></span>, <span class="highwire-citation-author" data-delta="12"><span class="nlm-given-names">Diane</span> <span class="nlm-surname">Dickel</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="13"><a href="http://orcid.org/0000-0002-9953-3637" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Aimee</span> <span class="nlm-surname">Zuniga</span></span>, <span class="highwire-citation-author" data-delta="14"><span class="nlm-given-names">Axel</span> <span class="nlm-surname">Visel</span></span>, <span class="highwire-citation-author hw-author-orcid-logo-wrapper" data-delta="15"><a href="http://orcid.org/0000-0002-3186-7403" target="_blank" class="hw-author-orcid-logo link-icon-only link-icon"><span class="hw-icon-orcid hw-icon-color-orcid"></span> <span class="title element-invisible">View ORCID Profile</span></a><span class="nlm-given-names">Rolf</span> <span class="nlm-surname">Zeller</span></span>, <span class="highwire-citation-author" data-delta="16"><span class="nlm-given-names">Licia</span> <span class="nlm-surname">Selleri</span></span></span></div> <div class="highwire-cite-metadata" ><span class="highwire-cite-metadata-doi highwire-cite-metadata"><span class="label">doi:</span> https://doi.org/10.1101/2022.03.08.483529 </span></div> </div> <div id="hw-article-author-popups-node-2437741--21842151150" style="display: none;"><div class="author-tooltip-0"><div class="author-tooltip-name">Marta Losa </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>^</span><span class='nlm-institution'>Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco</span>, San Francisco, California, <span class='nlm-country'>USA</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Marta%2BLosa%2B" 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Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco</span>, San Francisco, California, <span class='nlm-country'>USA</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Jean%2BDenis%2BBenazet%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Benazet%20JD&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link last"><a href="/search/author1%3AJean%2BDenis%2BBenazet%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this 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class='author-affiliation'><span class='nlm-sup'>^</span><span class='nlm-institution'>Program in Craniofacial Biology, Institute for Human Genetics, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Department of Orofacial Sciences and Department of Anatomy, University of California at San Francisco</span>, San Francisco, California, <span class='nlm-country'>USA</span></div></span></div><ul class="author-tooltip-find-more"><li class="author-tooltip-gs-link first"><a href="/lookup/google-scholar?link_type=googlescholar&gs_type=author&author%5B0%5D=Licia%2BSelleri%2B" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on Google Scholar</a></li><li class="author-tooltip-pubmed-link"><a href="/lookup/external-ref?access_num=Selleri%20L&link_type=AUTHORSEARCH" target="_blank" class="" data-icon-position="" data-hide-link-title="0">Find this author on PubMed</a></li><li class="author-site-search-link"><a 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abstract-view "><span class="highwire-journal-article-marker-start"></span><div class="section abstract" id="abstract-1"><h2 class="">ABSTRACT</h2><p id="p-4">During development cell fates are specified by tightly controlled gene expression programs. PBX TALE transcription factors control gene regulatory networks (GRN) that direct vertebrate tissue patterning and organ morphogenesis. How PBX1/2 proteins acquire context-specific functions, despite widespread embryonic expression of <em>Pbx1/2</em>, remains elusive. In mouse limb buds, mesenchymal-specific loss of PBX1/2 or of the transcriptional regulator HAND2 results in similar phenotypes, suggesting that PBX1/2- and HAND2-dependent programs converge to control limb development. To investigate this scenario, we combined tissue-specific and temporally-controlled mutagenesis with multi-omics approaches using the murine hindlimb bud as a model. We reconstructed a GRN collaboratively directed by PBX1/2 and HAND2, demonstrating that <em>Pbx1</em>-<em>Hand2</em> genetically interact <em>in vivo</em> during hindlimb patterning, with PBX1 concomitantly acting as an upstream regulator of <em>Hand2</em>. At organismal-level resolution the GRN is active within restricted subsets of posterior-proximal hindlimb mesenchymal cells, wherein <em>Pbx1/2</em> and <em>Hand2</em> are co-expressed with their target genes. Genome-wide profiling of PBX1 binding across multiple tissues further revealed that HAND2 selects a subset of PBX-bound regions to impart limb patterning functionality. This research elucidates mechanisms underlying limb bud-specific functions by PBX1/2, while informing general principles by which promiscuous transcription factors cooperate with select cofactors to instruct distinct developmental programs.</p></div><h3>Competing Interest Statement</h3><p id="p-5">The authors have declared no competing interest.</p><div class="section fn-group" id="fn-group-1"><h2>Footnotes</h2><ul><li class="fn-others" id="fn-2"><p id="p-2"><a class="rev-xref" href="#xref-fn-2-1">↵</a><span class="fn-label">[]</span> Co-senior authors</p></li></ul></div><span class="highwire-journal-article-marker-end"></span></div><span class="related-urls"></span></div></div> </div> </div> <div class="panel-separator"></div><div class="panel-pane pane-biorxiv-copyright" > <div class="pane-content"> <div class="field field-name-field-highwire-copyright field-type-text field-label-inline clearfix"><div class="field-label">Copyright </div><div class="field-items"><div class="field-item even">The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.<span class="license-type-none"> All rights reserved. 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