title: Serine/threonine phosphatase Stp1 mediates post-transcriptional regulation of hemolysin, autolysis, and virulence of group B Streptococcus.
abstract: Elucidating how serine/threonine phosphatases regulate kinase function and bacterial virulence is critical for our ability to combat these infections. Group B streptococci (GBS) are β-hemolytic Gram-positive bacteria that cause invasive infections in humans. To adapt to environmental changes, GBS encodes signaling mechanisms comprising two component systems and eukaryotic-like enzymes. We have previously described the importance of the serine/threonine kinase Stk1 to GBS pathogenesis. However, how the presence or absence of the cognate serine/threonine phosphatase Stp1 affects Stk1 function and GBS virulence is not known. Here, we show that GBS deficient only in Stp1 expression are markedly reduced for their ability to cause systemic infections, exhibit decreased β-hemolysin/cytolysin activity, and show increased sensitivity to autolysis. Although transcription of genes important for β-hemolysin/cytolysin expression and export is similar to the wild type (WT), 294 genes (excluding stp1) showed altered expression in the stp1 mutant and included autolysin genes. Furthermore, phosphopeptide enrichment analysis identified that 35 serine/threonine phosphopeptides, corresponding to 27 proteins, were unique to the stp1 mutant. This included phosphorylation of ATP synthase, DNA and RNA helicases, and proteins important for cell division and protein synthesis. Collectively, our results indicate that Stp1 is important for appropriate regulation of Stk1 function, hemolysin activity, autolysis, and GBS virulence.
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Kellie
authors.initials: K
authors.lastname: Burnside
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Lembo
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Maria Isabel
authors.initials: MI
authors.lastname: Harrell
authors.affiliation: Department of Biology and Center for Microbial Sciences, San Diego State University, San Diego, California 92182.
authors.forename: Michael
authors.initials: M
authors.lastname: Gurney
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Liang
authors.initials: L
authors.lastname: Xue
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Nguyen-Thao
authors.initials: NT
authors.lastname: BinhTran
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: James E
authors.initials: JE
authors.lastname: Connelly
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Kelsea A
authors.initials: KA
authors.lastname: Jewell
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Byron Z
authors.initials: BZ
authors.lastname: Schmidt
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101.
authors.forename: Melissa
authors.initials: M
authors.lastname: de Los Reyes
authors.affiliation: Department of Biochemistry, Purdue University, West Lafayette, Indiana 47907.
authors.forename: Weiguo Andy
authors.initials: WA
authors.lastname: Tao
authors.affiliation: Department of Biology and Center for Microbial Sciences, San Diego State University, San Diego, California 92182.
authors.forename: Kelly S
authors.initials: KS
authors.lastname: Doran
authors.affiliation: Department of Pediatric Infectious Diseases, University of Washington School of Medicine and Seattle Children's Hospital Research Institute, Seattle, Washington 98101. Electronic address: lakshmi.rajagopal@seattlechildrens.org.
authors.forename: Lakshmi
authors.initials: L
authors.lastname: Rajagopal
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015239:Arylsulfotransferase / Q000378:metabolism*; D001921:Brain / Q000378:metabolism; D005786:Gene Expression Regulation; D006460:Hemolysin Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008833:Microcirculation; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D012323:RNA Processing, Post-Transcriptional; D051381:Rats; D013292:Streptococcus agalactiae / Q000378:metabolism*; D014774:Virulence
chemical_list: D006460:Hemolysin Proteins; C474028:Stk1 protein, Streptococcus agalactiae; D017346:Protein Serine-Threonine Kinases; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human; C573795:Sult1a1 protein, rat; D010749:Phosphoprotein Phosphatases
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