{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:55:49Z","timestamp":1768510549480,"version":"3.49.0"},"posted":{"date-parts":[[2024,4,8]]},"group-title":"Microbiology","reference-count":73,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,4,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The intracellular human pathogen\n                  <jats:italic>Shigella<\/jats:italic>\n                  invades the colonic epithelium to cause disease. Prior to invasion, this bacterium navigates through different environments within the human body, including the stomach and the small intestine. To adapt to changing environments,\n                  <jats:italic>Shigella<\/jats:italic>\n                  uses the bacterial second messenger c-di-GMP signaling system, synthesized by diguanylate cyclases (DGCs) encoding GGDEF domains.\n                  <jats:italic>Shigella flexneri<\/jats:italic>\n                  encodes a total of 9 GGDEF or GGDEF-EAL domain enzymes in its genome, but 5 of these genes have acquired mutations that presumably inactivated the c-di-GMP synthesis activity of these enzymes. In this study, we examined individual\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  DGCs for their role in c-di-GMP synthesis and pathogenesis. We individually expressed each of the 4 intact DGCs in an\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  strain where these 4 DGCs had been deleted (\u03944DGC). We found that the 4\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  intact DGCs synthesize c-di-GMP at different levels\n                  <jats:italic>in vitro<\/jats:italic>\n                  and during infection of tissue-cultured cells. We also found that\n                  <jats:italic>dgcF<\/jats:italic>\n                  and\n                  <jats:italic>dgcI<\/jats:italic>\n                  expression significantly reduces invasion and plaque formation, and\n                  <jats:italic>dgcF<\/jats:italic>\n                  expression increases acid sensitivity, and that these phenotypes did not correspond with measured c-di-GMP levels. However, deletion of these 4 DGCs did not eliminate\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  c-di-GMP, and we found that\n                  <jats:italic>dgcE, dgcQ,<\/jats:italic>\n                  and\n                  <jats:italic>dgcN<\/jats:italic>\n                  , which all have nonsense mutations prior to the GGDEF domain, still produce c-di-GMP. These\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  degenerate DGC genes are expressed as multiple proteins, consistent with multiple start codons within the gene. We propose that both intact and degenerate DGCs contribute to\n                  <jats:italic>S. flexneri<\/jats:italic>\n                  c-di-GMP signaling.\n                <\/jats:p>","DOI":"10.1101\/2024.04.08.588579","type":"posted-content","created":{"date-parts":[[2024,4,9]],"date-time":"2024-04-09T01:10:21Z","timestamp":1712625021000},"source":"Crossref","is-referenced-by-count":0,"title":["Intact and Degenerate Diguanylate Cyclases regulate\n                  <i>Shigella<\/i>\n                  Cyclic di-GMP"],"prefix":"10.64898","author":[{"given":"Ruchi","family":"Ojha","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefanie","family":"Krug","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Prentiss","family":"Jones","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7213-0953","authenticated-orcid":false,"given":"Benjamin J.","family":"Koestler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024041012200606000_2024.04.08.588579v1.1","first-page":"651","article-title":"Global burden of Shigella infections: implications for vaccine development and implementation of control strategies","volume":"77","year":"1999","journal-title":"Bull World Health Organ"},{"key":"2024041012200606000_2024.04.08.588579v1.2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/13.Supplement_4.S245"},{"key":"2024041012200606000_2024.04.08.588579v1.3","doi-asserted-by":"crossref","first-page":"2474","DOI":"10.1128\/iai.64.7.2474-2482.1996","article-title":"Analysis of epithelial cell stress response during infection by Shigella flexneri","volume":"64","year":"1996","journal-title":"Infect Immun"},{"key":"2024041012200606000_2024.04.08.588579v1.4","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.43.1.195-201.1984"},{"key":"2024041012200606000_2024.04.08.588579v1.5","unstructured":"CDC. 2013. 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