title: Mutual exclusivity of hyaluronan and hyaluronidase in invasive group A Streptococcus.
abstract: A recent analysis of group A Streptococcus (GAS) invasive infections in Australia has shown a predominance of M4 GAS, a serotype recently reported to lack the antiphagocytic hyaluronic acid (HA) capsule. Here, we use molecular genetics and bioinformatics techniques to characterize 17 clinical M4 isolates associated with invasive disease in children during this recent epidemiology. All M4 isolates lacked HA capsule, and whole genome sequence analysis of two isolates revealed the complete absence of the hasABC capsule biosynthesis operon. Conversely, M4 isolates possess a functional HA-degrading hyaluronate lyase (HylA) enzyme that is rendered nonfunctional in other GAS through a point mutation. Transformation with a plasmid expressing hasABC restored partial encapsulation in wild-type (WT) M4 GAS, and full encapsulation in an isogenic M4 mutant lacking HylA. However, partial encapsulation reduced binding to human complement regulatory protein C4BP, did not enhance survival in whole human blood, and did not increase virulence of WT M4 GAS in a mouse model of systemic infection. Bioinformatics analysis found no hasABC homologs in closely related species, suggesting that this operon was a recent acquisition. These data showcase a mutually exclusive interaction of HA capsule and active HylA among strains of this leading human pathogen.
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093; School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Anna
authors.initials: A
authors.lastname: Henningham
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093; Department of Oral and Molecular Microbiology, Osaka University Graduate School of Dentistry, Suita, Osaka 565-0871, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Yamaguchi
authors.affiliation: Systems Biology Research Group, University of California San Diego, La Jolla, California 92093; Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo 11562, Egypt.
authors.forename: Ramy K
authors.initials: RK
authors.lastname: Aziz
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093; Department of Pediatrics, Laboratory of Pediatric Infectious Diseases, Radboud University Medical Centre, 6500 HC Nijmegen, The Netherlands.
authors.forename: Kirsten
authors.initials: K
authors.lastname: Kuipers
authors.affiliation: Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, California 92093.
authors.forename: Cosmo Z
authors.initials: CZ
authors.lastname: Buffalo
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093.
authors.forename: Samira
authors.initials: S
authors.lastname: Dahesh
authors.affiliation: Glycobiology Research and Training Center, University of California San Diego, La Jolla, California 92093.
authors.forename: Biswa
authors.initials: B
authors.lastname: Choudhury
authors.affiliation: Glycobiology Research and Training Center, University of California San Diego, La Jolla, California 92093.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Van Vleet
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Yamaguchi
authors.affiliation: School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Seymour
authors.affiliation: School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Nouri L
authors.initials: NL
authors.lastname: Ben Zakour
authors.affiliation: Department of Mathematics and Statistics, San Diego State University, San Diego, California 92182.
authors.forename: Lingjun
authors.initials: L
authors.lastname: He
authors.affiliation: Queensland Health Forensic and Scientific Services, Coopers Plains, Queensland 4108, Australia.
authors.forename: Helen V
authors.initials: HV
authors.lastname: Smith
authors.affiliation: Queensland Children's Medical Research Institute, Herston, Queensland 4029, Australia, and.
authors.forename: Keith
authors.initials: K
authors.lastname: Grimwood
authors.affiliation: School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Beatson
authors.affiliation: Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, California 92093.
authors.forename: Partho
authors.initials: P
authors.lastname: Ghosh
authors.affiliation: School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Walker
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093; Skaggs School of Pharmacy and Pharmaceutical Sciences, and University of California San Diego, La Jolla, California 92093; Rady Children's Hospital, San Diego, California 92123.
authors.forename: Victor
authors.initials: V
authors.lastname: Nizet
authors.affiliation: Department of Pediatrics, University of California San Diego, La Jolla, California 92093; School of Chemistry and Molecular Biosciences and The University of Queensland, St. Lucia, Queensland 4072, Australia; Australian Infectious Diseases Research Centre, The University of Queensland, St. Lucia, Queensland 4072, Australia,. Electronic address: jncole@ucsd.edu.
authors.forename: Jason N
authors.initials: JN
authors.lastname: Cole
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001483:Base Sequence; D002462:Cell Membrane / Q000382:microbiology; D019295:Computational Biology; D005098:Exotoxins / Q000378:metabolism; D005260:Female; D005816:Genetic Complementation Test; D000071677:Histidine Kinase; D006801:Humans; D006820:Hyaluronic Acid / Q000378:metabolism*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000382:microbiology; D017354:Point Mutation; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D013290:Streptococcal Infections / Q000382:microbiology*; D013297:Streptococcus pyogenes / Q000201:enzymology*; D014774:Virulence
chemical_list: D001426:Bacterial Proteins; C432143:CsrR protein, Streptococcus pyogenes; D005098:Exotoxins; D047908:Intracellular Signaling Peptides and Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D012097:Repressor Proteins; C098249:RopB protein, Rhizobium leguminosarum; C027951:erythrogenic toxin; D006820:Hyaluronic Acid; C534547:CovS protein, Streptococcus pyogenes; D000071677:Histidine Kinase; D006821:Hyaluronoglucosaminidase; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase
keywords: Bacterial Pathogenesis; Group A Streptococcus; Hyaluronan; Hyaluronate; Hyaluronate Lyase; Hyaluronic acid Capsule; Infectious Disease; Invasive Disease; Nonencapsulated; Streptococcus Pyogenes (S. Pyogenes)