title: Analysis of polymorphic residues reveals distinct enzymatic and cytotoxic activities of the Streptococcus pyogenes NAD+ glycohydrolase.
abstract: The Streptococcus pyogenes NAD(+) glycohydrolase (SPN) is secreted from the bacterial cell and translocated into the host cell cytosol where it contributes to cell death. Recent studies suggest that SPN is evolving and has diverged into NAD(+) glycohydrolase-inactive variants that correlate with tissue tropism. However, the role of SPN in both cytotoxicity and niche selection are unknown. To gain insight into the forces driving the adaptation of SPN, a detailed comparison of representative glycohydrolase activity-proficient and -deficient variants was conducted. Of a total 454 amino acids, the activity-deficient variants differed at only nine highly conserved positions. Exchanging residues between variants revealed that no one single residue could account for the inability of the deficient variants to cleave the glycosidic bond of β-NAD(+) into nicotinamide and ADP-ribose; rather, reciprocal changes at 3 specific residues were required to both abolish activity of the proficient version and restore full activity to the deficient variant. Changing any combination of 1 or 2 residues resulted in intermediate activity. However, a change to any 1 residue resulted in a significant decrease in enzyme efficiency. A similar pattern involving multiple residues was observed for comparison with a second highly conserved activity-deficient variant class. Remarkably, despite differences in glycohydrolase activity, all versions of SPN were equally cytotoxic to cultured epithelial cells. These data indicate that the glycohydrolase activity of SPN may not be the only contribution the toxin has to the pathogenesis of S. pyogenes and that both versions of SPN play an important role during infection.
authors.affiliation: Department of Molecular Microbiology Washington University School of Medicine, St Louis, Missouri 63110-1093, USA.
authors.forename: Sukantha
authors.initials: S
authors.lastname: Chandrasekaran
authors.forename: Joydeep
authors.initials: J
authors.lastname: Ghosh
authors.forename: Gary C
authors.initials: GC
authors.lastname: Port
authors.forename: Eun-Ik
authors.initials: EI
authors.lastname: Koh
authors.forename: Michael G
authors.initials: MG
authors.lastname: Caparon
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004847:Epithelial Cells / Q000201:enzymology* / Q000382:microbiology / Q000473:pathology; D006801:Humans; D009243:NAD / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009244:NAD+ Nucleosidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D013290:Streptococcal Infections / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D013297:Streptococcus pyogenes / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D009243:NAD; D000244:Adenosine Diphosphate; D009244:NAD+ Nucleosidase
keywords: Allelic Variation; Bacteria; Bacterial Pathogenesis; Bacterial Toxins; Cell Death; Cytotoxicity; Enzymes; Group A Streptococcus; NAD+ Glycohydrolase