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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="Natural transformation (NT) is a conserved mechanism of horizontal gene transfer in bacterial species. During this process, DNA is taken up into the cytoplasm where it can be integrated into the host genome by homologous recombination. We have previously shown that some cytoplasmic exonucleases inhibit NT by degrading ingested DNA prior to its successful recombination. However, one exonuclease, SbcB, counterintuitively promotes NT in Vibrio cholerae . Here, through a systematic analysis of the distinct steps of NT, we show that SbcB acts downstream of DNA uptake into the cytoplasm, but upstream of recombinational branch migration. Through mutational analysis, we show that SbcB promotes NT in a manner that does not rely on its exonuclease activity. Finally, we provide genetic evidence that SbcB directly interacts with the primary bacterial recombinase, RecA. Together, these data advance our molecular understanding of horizontal gene transfer in V. cholerae , and reveal that SbcB promotes homologous recombination during NT in a manner that does not rely on its canonical exonuclease activity. IMPORTANCE Horizontal gene transfer by natural transformation contributes to the spread of antibiotic resistance and virulence factors in bacterial species. Here, we study how one protein, SbcB, helps facilitate this process in the facultative bacterial pathogen Vibrio cholerae . SbcB is a well-known for its exonuclease activity ( i . e ., the ability to degrade the ends of linear DNA). Through this study we uncover that while SbcB is important for natural transformation, it does not facilitate this process using its exonuclease activity. Thus, this work helps further our understanding of the molecular events required for this conserved evolutionary process, and uncovers a function for SbcB beyond its canonical exonuclease activity. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="DC.Contributor" content="Triana N. Dalia" /> <meta name="DC.Contributor" content="Ankur B. Dalia" /> <meta name="article:published_time" content="2024-11-18" /> <meta name="article:section" content="New Results" /> <meta name="citation_title" content="SbcB facilitates natural transformation in Vibrio cholerae in an exonuclease-independent manner" /> <meta name="citation_abstract" lang="en" content="<p>Natural transformation (NT) is a conserved mechanism of horizontal gene transfer in bacterial species. During this process, DNA is taken up into the cytoplasm where it can be integrated into the host genome by homologous recombination. We have previously shown that some cytoplasmic exonucleases inhibit NT by degrading ingested DNA prior to its successful recombination. However, one exonuclease, SbcB, counterintuitively promotes NT in Vibrio cholerae. Here, through a systematic analysis of the distinct steps of NT, we show that SbcB acts downstream of DNA uptake into the cytoplasm, but upstream of recombinational branch migration. Through mutational analysis, we show that SbcB promotes NT in a manner that does not rely on its exonuclease activity. Finally, we provide genetic evidence that SbcB directly interacts with the primary bacterial recombinase, RecA. Together, these data advance our molecular understanding of horizontal gene transfer in V. cholerae, and reveal that SbcB promotes homologous recombination during NT in a manner that does not rely on its canonical exonuclease activity.</p>" /> <meta name="citation_journal_title" content="bioRxiv" /> <meta name="citation_publisher" content="Cold Spring Harbor Laboratory" /> <meta name="citation_publication_date" content="2024/01/01" /> <meta name="citation_mjid" content="biorxiv;2024.09.25.615017v2" /> <meta name="citation_id" content="2024.09.25.615017v2" /> <meta name="citation_public_url" content="https://www.biorxiv.org/content/10.1101/2024.09.25.615017v2" /> <meta name="citation_abstract_html_url" content="https://www.biorxiv.org/content/10.1101/2024.09.25.615017v2.abstract" /> <meta name="citation_full_html_url" content="https://www.biorxiv.org/content/10.1101/2024.09.25.615017v2.full" /> <meta name="citation_pdf_url" content="https://www.biorxiv.org/content/biorxiv/early/2024/11/18/2024.09.25.615017.full.pdf" /> <meta name="citation_doi" content="10.1101/2024.09.25.615017" /> <meta name="citation_num_pages" content="23" /> <meta name="citation_article_type" content="Article" /> <meta name="citation_section" content="New Results" /> <meta name="citation_firstpage" content="2024.09.25.615017" /> <meta name="citation_author" content="Triana N. Dalia" /> <meta name="citation_author_institution" content="Department of Biology" /> <meta name="citation_author" content="Ankur B. Dalia" /> <meta name="citation_author_institution" content="Department of Biology" /> <meta name="citation_author_email" content="ankdalia@iu.edu" /> <meta name="citation_reference" content="Ellison CK, Dalia TN, Vidal Ceballos A, Wang JC, Biais N, Brun YV, Dalia AB. 2018. Retraction of DNA-bound type IV competence pili initiates DNA uptake during natural transformation in Vibrio cholerae. Nat Microbiol 3:773–780." /> <meta name="citation_reference" content="Dubnau D, Blokesch M. 2019. Mechanisms of DNA Uptake by Naturally Competent Bacteria. Annu Rev Genet 53:217–237." /> <meta name="citation_reference" content="Gangel H, Hepp C, Muller S, Oldewurtel ER, Aas FE, Koomey M, Maier B. 2014. Concerted spatio-temporal dynamics of imported DNA and ComE DNA uptake protein during gonococcal transformation. 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J Bacteriol 177:4121–30." /> <meta name="twitter:title" content="SbcB facilitates natural transformation in Vibrio cholerae in an exonuclease-independent manner" /> <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="Natural transformation (NT) is a conserved mechanism of horizontal gene transfer in bacterial species. During this process, DNA is taken up into the cytoplasm where it can be integrated into the host genome by homologous recombination. We have previously shown that some cytoplasmic exonucleases inhibit NT by degrading ingested DNA prior to its successful recombination. However, one exonuclease, SbcB, counterintuitively promotes NT in Vibrio cholerae . Here, through a systematic analysis of the distinct steps of NT, we show that SbcB acts downstream of DNA uptake into the cytoplasm, but upstream of recombinational branch migration. Through mutational analysis, we show that SbcB promotes NT in a manner that does not rely on its exonuclease activity. Finally, we provide genetic evidence that SbcB directly interacts with the primary bacterial recombinase, RecA. Together, these data advance our molecular understanding of horizontal gene transfer in V. cholerae , and reveal that SbcB promotes homologous recombination during NT in a manner that does not rely on its canonical exonuclease activity. IMPORTANCE Horizontal gene transfer by natural transformation contributes to the spread of antibiotic resistance and virulence factors in bacterial species. Here, we study how one protein, SbcB, helps facilitate this process in the facultative bacterial pathogen Vibrio cholerae . SbcB is a well-known for its exonuclease activity ( i . e ., the ability to degrade the ends of linear DNA). Through this study we uncover that while SbcB is important for natural transformation, it does not facilitate this process using its exonuclease activity. Thus, this work helps further our understanding of the molecular events required for this conserved evolutionary process, and uncovers a function for SbcB beyond its canonical exonuclease activity. ### Competing Interest Statement The authors have declared no competing interest." /> <meta name="og-title" property="og:title" content="SbcB facilitates natural transformation in Vibrio cholerae in an exonuclease-independent manner" /> <meta name="og-url" property="og:url" content="https://www.biorxiv.org/content/10.1101/2024.09.25.615017v2" /> <meta name="og-site-name" property="og:site_name" content="bioRxiv" /> <meta name="og-description" property="og:description" content="Natural transformation (NT) is a conserved mechanism of horizontal gene transfer in bacterial species. During this process, DNA is taken up into the cytoplasm where it can be integrated into the host genome by homologous recombination. We have previously shown that some cytoplasmic exonucleases inhibit NT by degrading ingested DNA prior to its successful recombination. However, one exonuclease, SbcB, counterintuitively promotes NT in Vibrio cholerae . Here, through a systematic analysis of the distinct steps of NT, we show that SbcB acts downstream of DNA uptake into the cytoplasm, but upstream of recombinational branch migration. Through mutational analysis, we show that SbcB promotes NT in a manner that does not rely on its exonuclease activity. Finally, we provide genetic evidence that SbcB directly interacts with the primary bacterial recombinase, RecA. Together, these data advance our molecular understanding of horizontal gene transfer in V. cholerae , and reveal that SbcB promotes homologous recombination during NT in a manner that does not rely on its canonical exonuclease activity. IMPORTANCE Horizontal gene transfer by natural transformation contributes to the spread of antibiotic resistance and virulence factors in bacterial species. Here, we study how one protein, SbcB, helps facilitate this process in the facultative bacterial pathogen Vibrio cholerae . SbcB is a well-known for its exonuclease activity ( i . e ., the ability to degrade the ends of linear DNA). Through this study we uncover that while SbcB is important for natural transformation, it does not facilitate this process using its exonuclease activity. Thus, this work helps further our understanding of the molecular events required for this conserved evolutionary process, and uncovers a function for SbcB beyond its canonical exonuclease activity. ### 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="2024-11-18" /> <link rel="alternate" type="application/vnd.ms-powerpoint" title="Powerpoint" href="/content/10.1101/2024.09.25.615017v2.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" href="https://www.biorxiv.org/content/10.1101/2024.09.25.615017v2" /> <link 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class="highwire-article-citation highwire-citation-type-highwire-article node4230966" data-node-nid="4230966" id="node-4230966--2338278236" data-pisa="biorxiv;2024.09.25.615017v2" data-pisa-master="biorxiv;2024.09.25.615017" data-apath="/biorxiv/early/2024/11/18/2024.09.25.615017.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">SbcB facilitates natural transformation in <em>Vibrio cholerae</em> in an exonuclease-independent manner</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">Triana N.</span> <span class="nlm-surname">Dalia</span></span>, <span class="highwire-citation-author" data-delta="1"><span class="nlm-given-names">Ankur B.</span> <span class="nlm-surname">Dalia</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/10.1101/2024.09.25.615017 </span></div> </div> <div id="hw-article-author-popups-node-4230966--2338278236" style="display: none;"><div class="author-tooltip-0"><div class="author-tooltip-name">Triana N. Dalia </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>1</span><span class='nlm-institution'>Department of Biology</span>, Indiana University, Bloomington, IN 47405</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=Triana%2BN.%2BDalia%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=Dalia%20TN&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%3ATriana%2BN.%2BDalia%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this site</a></li></ul></div><div class="author-tooltip-1"><div class="author-tooltip-name">Ankur B. Dalia </div><div class="author-tooltip-affiliation"><span class="author-tooltip-text"><div class='author-affiliation'><span class='nlm-sup'>1</span><span class='nlm-institution'>Department of Biology</span>, Indiana University, Bloomington, IN 47405</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=Ankur%2BB.%2BDalia%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=Dalia%20AB&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 href="/search/author1%3AAnkur%2BB.%2BDalia%2B" rel="nofollow" class="" data-icon-position="" data-hide-link-title="0">Search for this author on this site</a></li><li class="author-corresp-email-link last"><span>For correspondence: <a href="mailto:ankdalia@iu.edu" class="" data-icon-position="" data-hide-link-title="0">ankdalia@iu.edu</a></span></li></ul></div></div></div> </div> </div> <div class="panel-separator"></div><div class="panel-pane pane-highwire-panel-tabs pane-panels-ajax-tab-tabs" > <div class="pane-content"> <div class="item-list"><ul class="tabs inline panels-ajax-tab"><li class="first"><a href="/content/10.1101/2024.09.25.615017v2" class="panels-ajax-tab-tab" data-panel-name="biorxiv_tab_art" data-target-id="highwire_article_tabs" data-entity-context="node:4230966" data-trigger="" data-url-enabled="1">Abstract</a><a href="/panels_ajax_tab/biorxiv_tab_art/node:4230966/1" rel="nofollow" style="display:none" class="js-crawler-link"></a></li><li><a href="/content/10.1101/2024.09.25.615017v2.full-text" class="panels-ajax-tab-tab" data-panel-name="article_tab_full_text" data-target-id="highwire_article_tabs" 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pane-highwire-panel-tabs-container" > <div class="pane-content"> <div data-panels-ajax-tab-preloaded="biorxiv_tab_art" id="panels-ajax-tab-container-highwire_article_tabs" class="panels-ajax-tab-container"><div class="panels-ajax-tab-loading" style ="display:none"><img class="loading" src="https://www.biorxiv.org/sites/all/modules/contrib/panels_ajax_tab/images/loading.gif" alt="Loading" title="Loading" /></div><div class="panels-ajax-tab-wrap-biorxiv_tab_art"><div class="panel-display panel-1col clearfix" > <div class="panel-panel panel-col"> <div><div class="panel-pane pane-highwire-markup" > <div class="pane-content"> <div class="highwire-markup"><div xmlns="http://www.w3.org/1999/xhtml" data-highwire-cite-ref-tooltip-instance="highwire_reflinks_tooltip" class="content-block-markup" xmlns:xhtml="http://www.w3.org/1999/xhtml"><div class="article abstract-view "><span class="highwire-journal-article-marker-start"></span><div class="section abstract" id="abstract-1"><h2 class="">ABSTRACT</h2><p id="p-2">Natural transformation (NT) is a conserved mechanism of horizontal gene transfer in bacterial species. During this process, DNA is taken up into the cytoplasm where it can be integrated into the host genome by homologous recombination. We have previously shown that some cytoplasmic exonucleases inhibit NT by degrading ingested DNA prior to its successful recombination. However, one exonuclease, SbcB, counterintuitively promotes NT in <em>Vibrio cholerae</em>. Here, through a systematic analysis of the distinct steps of NT, we show that SbcB acts downstream of DNA uptake into the cytoplasm, but upstream of recombinational branch migration. Through mutational analysis, we show that SbcB promotes NT in a manner that does not rely on its exonuclease activity. Finally, we provide genetic evidence that SbcB directly interacts with the primary bacterial recombinase, RecA. Together, these data advance our molecular understanding of horizontal gene transfer in <em>V. cholerae</em>, and reveal that SbcB promotes homologous recombination during NT in a manner that does not rely on its canonical exonuclease activity.</p><div id="sec-1" class="subsection"><p id="p-3"><strong>IMPORTANCE</strong> Horizontal gene transfer by natural transformation contributes to the spread of antibiotic resistance and virulence factors in bacterial species. Here, we study how one protein, SbcB, helps facilitate this process in the facultative bacterial pathogen <em>Vibrio cholerae</em>. SbcB is a well-known for its exonuclease activity (<em>i</em>.<em>e</em>., the ability to degrade the ends of linear DNA). Through this study we uncover that while SbcB is important for natural transformation, it does not facilitate this process using its exonuclease activity. Thus, this work helps further our understanding of the molecular events required for this conserved evolutionary process, and uncovers a function for SbcB beyond its canonical exonuclease activity.</p></div></div><h3>Competing Interest Statement</h3><p id="p-4">The authors have declared no competing interest.</p><div class="section fn-group" id="fn-group-1"><h2>Footnotes</h2><ul><li class="fn-update fn-group-summary-of-updates" id="fn-1"><p id="p-5">Performed new experiments and adjusted the figures (Fig. 5 and Fig. 6C) and text accordingly.</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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