{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:07:49Z","timestamp":1785434869789,"version":"3.56.0"},"posted":{"date-parts":[[2024,11,5]]},"group-title":"Microbiology","reference-count":36,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,5]],"date-time":"2024-11-05T00:00:00Z","timestamp":1730764800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["09150181910001"],"award-info":[{"award-number":["09150181910001"]}],"id":[{"id":"https:\/\/ror.org\/01yaj9a77","id-type":"ROR","asserted-by":"publisher"}]},{"name":"NIH-NIAID","award":["R01 AI143690"],"award-info":[{"award-number":["R01 AI143690"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,5,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  <jats:italic>Streptococcus pyogenes<\/jats:italic>\n                  (\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  ) is a leading cause of infection-related mortality in humans globally. The characteristic cell wall-anchored Group A Carbohydrate (GAC) is expressed by all\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  strains and consists of a polyrhamnose backbone with alternating\n                  <jats:italic>N<\/jats:italic>\n                  -acetylglucosamine (GlcNAc) side chains, of which 25% are decorated with glycerol phosphate (GroP). The genes in the\n                  <jats:italic>gacA-L<\/jats:italic>\n                  cluster are critical for GAC biosynthesis with\n                  <jats:italic>gacH-L<\/jats:italic>\n                  being responsible for the characteristic GlcNAc-GroP decoration, which confers the agglutination in rapid test diagnostic assays and contributes to\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  pathogenicity. Historical research papers described\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  isolates, so-called A-variant strains, that lost the characteristic GlcNAc side chain following serial animal passage. Genomic analysis of a single viable historic parent\/A-variant strain pair revealed a premature inactivating stop codon in\n                  <jats:italic>gacI<\/jats:italic>\n                  , explaining the described loss of the GlcNAc side chain. Subsequently, we analyzed the genetic variation of the 12\n                  <jats:italic>gacA-L<\/jats:italic>\n                  genes in a collection of 2,021\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  genome sequences. Although all\n                  <jats:italic>gac<\/jats:italic>\n                  genes (\n                  <jats:italic>gacA-L<\/jats:italic>\n                  ) displayed genetic variation, we only identified 26 isolates (1.3%) with a premature stop codon in one of the\n                  <jats:italic>gac<\/jats:italic>\n                  genes. Twelve out of 26 (46%) isolates contained a premature stop codon in\n                  <jats:italic>gacH<\/jats:italic>\n                  , which encodes the enzyme responsible for the GroP modification. To study the functional consequences of the different premature stop codons for GacH function, we plasmid-expressed three\n                  <jats:italic>gacH<\/jats:italic>\n                  variants in a\n                  <jats:italic>S. pyogenes gacH<\/jats:italic>\n                  -deficient strain. Cell wall analysis confirmed GacH loss-of-function for the studied\n                  <jats:italic>gacH<\/jats:italic>\n                  variants through the significant reduction of GAC GroP, complete resistance to killing by the human bactericidal enzyme group IIA-secreted phospholipase, and susceptibility to zinc toxicity. Overall, our data provide a comprehensive overview of the genetic variation of the\n                  <jats:italic>gacA-L<\/jats:italic>\n                  cluster in a global population of\n                  <jats:italic>S. pyogenes<\/jats:italic>\n                  strains and the functional consequences of rare inactivating mutations in\n                  <jats:italic>gacH<\/jats:italic>\n                  for host interaction.\n                <\/jats:p>","DOI":"10.1101\/2024.11.04.621835","type":"posted-content","created":{"date-parts":[[2024,11,6]],"date-time":"2024-11-06T04:36:04Z","timestamp":1730867764000},"source":"Crossref","is-referenced-by-count":1,"title":["Natural variation of the streptococcal Group A carbohydrate biosynthesis genes impacts host-pathogen interaction"],"prefix":"10.64898","author":[{"given":"Kim","family":"Schipper","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6194-2400","authenticated-orcid":false,"given":"Sara M.","family":"Tamminga","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-7263-7193","authenticated-orcid":false,"given":"Nicholas","family":"Murner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9752-6273","authenticated-orcid":false,"given":"Matthew","family":"Davies","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1129-1650","authenticated-orcid":false,"given":"Paul","family":"Berkhout","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8061-7750","authenticated-orcid":false,"given":"Debra E.","family":"Bessen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-9014-0014","authenticated-orcid":false,"given":"Astrid","family":"Hendriks","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8696-4892","authenticated-orcid":false,"given":"Natalia","family":"Korotkova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1154-652X","authenticated-orcid":false,"given":"Yvonne","family":"Pannekoek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2695-5863","authenticated-orcid":false,"given":"Nina M.","family":"van Sorge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025060300050513000_2024.11.04.621835v2.1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.6.1822"},{"key":"2025060300050513000_2024.11.04.621835v2.2","unstructured":"Amy Sims Sanyahumbi SC , Rosemary Wyber , and Jonathan R. 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