{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,8,2]],"date-time":"2025-08-02T14:43:46Z","timestamp":1754145826501,"version":"3.41.2"},"reference-count":35,"publisher":"Microbiology Society","issue":"7","license":[{"start":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T00:00:00Z","timestamp":1752710400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ZonMw","award":["09150181910001"],"award-info":[{"award-number":["09150181910001"]}]},{"DOI":"10.13039\/100006492","name":"Division of Intramural Research, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"crossref","award":["AI143690"],"award-info":[{"award-number":["AI143690"]}],"id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100010269","name":"Wellcome Trust","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100010269","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,7,17]]},"abstract":"<jats:p>\n                  <jats:italic>Streptococcus pyogenes<\/jats:italic> is a leading cause of infection-related mortality in humans globally. The characteristic cell wall-anchored group A carbohydrate (GAC) is expressed by all <jats:italic>S. pyogenes<\/jats:italic> strains and consists of a polyrhamnose backbone with alternating <jats:italic>N<\/jats:italic>-acetylglucosamine (GlcNAc) side chains, of which 25% are decorated with glycerol phosphate (GroP). The genes in the <jats:italic>gacA-L<\/jats:italic> cluster are critical for GAC biosynthesis, with <jats:italic>gacH-L<\/jats:italic> being responsible for the characteristic GlcNAc\u2013GroP decoration, which confers the agglutination in rapid test diagnostic assays and contributes to <jats:italic>S. pyogenes<\/jats:italic> pathogenicity. Historical research papers described <jats:italic>S. pyogenes<\/jats:italic> 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 <jats:italic>gacI<\/jats:italic>, explaining the described loss of the GlcNAc side chain. Subsequently, we analysed the genetic variation of the 12 <jats:italic>gacA-L<\/jats:italic> genes in a collection of 2021\u2009<jats:italic>S<\/jats:italic>. <jats:italic>pyogenes<\/jats:italic> genome sequences. Although all <jats:italic>gac<\/jats:italic> genes (<jats:italic>gacA-L<\/jats:italic>) displayed genetic variation, we only identified 26 isolates (1.3%) with a premature stop codon in one of the <jats:italic>gac<\/jats:italic> genes. Twelve out of 26 (46%) isolates contained a premature stop codon in <jats:italic>gacH<\/jats:italic>, 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 <jats:italic>gacH<\/jats:italic> variants in a <jats:italic>S. pyogenes gacH<\/jats:italic>-deficient strain. Cell wall analysis confirmed GacH loss of function for the studied <jats:italic>gacH<\/jats:italic> 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 <jats:italic>gacA-L<\/jats:italic> cluster in a global population of <jats:italic>S. pyogenes<\/jats:italic> strains and the functional consequences of rare inactivating mutations in <jats:italic>gacH<\/jats:italic> for host interaction.<\/jats:p>","DOI":"10.1099\/mgen.0.001443","type":"journal-article","created":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T13:17:43Z","timestamp":1752758263000},"source":"Crossref","is-referenced-by-count":0,"title":["Natural variation of the streptococcal group A carbohydrate biosynthesis genes impacts host\u2013pathogen interaction"],"prefix":"10.1099","volume":"11","author":[{"given":"Kim","family":"Schipper","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology and Infection Prevention, Amsterdam University Medical Center, University of Amsterdam, Amsterdam, 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