Characterization of a Serine/Threonine Kinase Involved in Virulence of Staphylococcus aureus

被引:105
作者
Debarbouille, Michel [1 ,2 ]
Dramsi, Shaynoor [1 ,2 ]
Dussurget, Olivier [3 ,4 ,5 ]
Nahori, Marie-Anne [3 ,4 ,5 ]
Vaganay, Elisabeth [6 ]
Jouvion, Gregory [7 ]
Cozzone, Alain [6 ]
Msadek, Tarek [1 ,2 ]
Duclos, Bertrand [6 ]
机构
[1] Inst Pasteur, Unite Biol Bacteries Pathogenes Gram Positif, F-75724 Paris 15, France
[2] CNRS, URA 2172, F-75724 Paris 15, France
[3] Inst Pasteur, Unite Interact Bacteries Cellules, F-75724 Paris 15, France
[4] INSERM, U604, F-75724 Paris 15, France
[5] INRA, USC2020, F-75724 Paris 15, France
[6] Univ Lyon, CNRS, Inst Biol & Chim Prot, Unite Mixte Rech 5086, Lyon, France
[7] Inst Pasteur, Unite Rech & Expertise Histotechnol & Pathol, F-75724 Paris 15, France
关键词
SERINE-THREONINE PHOSPHATASE; GRAM-POSITIVE BACTERIA; BACILLUS-SUBTILIS; PROTEIN-KINASE; STREPTOCOCCUS-PNEUMONIAE; SER/THR KINASE; BIOCHEMICAL-CHARACTERIZATION; PHYSIOLOGICAL ROLES; SIGNAL-TRANSDUCTION; MYXOCOCCUS-XANTHUS;
D O I
10.1128/JB.01813-08
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Staphylococcus aureus is a common human cutaneous and nasal commensal and a major life-threatening pathogen. Adaptation to the different environments encountered inside and outside the host is a crucial requirement for survival and colonization. We identified and characterized a eukaryotic-like serine/threonine kinase with three predicted extracellular PASTA domains (SA1063, or Stk1) and its associated phosphatase (SA1062, or Stp1) in S. aureus. Biochemical analyses revealed that Stk1 displays autokinase activity on threonine and serine residues and is localized to the membrane. Stp1 is a cytoplasmic protein with manganese-dependent phosphatase activity toward phosphorylated Stk1. In-frame deletions of the stk1 and stp1 genes were constructed in S. aureus strain 8325-4. Phenotypic analyses of the mutants revealed reduced growth of the stk1 mutant in RPMI 1640 defined medium that was restored when adenine was added to the medium. Furthermore, the stk1 mutant displayed increased resistance to Triton X-100 and to fosfomycin, suggesting modifications in cell wall metabolism. The stk1 mutant was tested for virulence in a mouse pyelonephritis model and found to be strongly reduced for survival in the kidneys (approximately 2-log-unit decrease) compared to the parental strain. Renal histopathological analyses showed severe inflammatory lesions in mice infected with the parental S. aureus SH1000 strain, whereas the Delta stk1 mutant led to only minimal renal lesions. These results confirm the important role of Stk1 for full expression of S. aureus pathogenesis and suggest that phosphorylation levels controlled by stk1 are essential in controlling bacterial survival within the host.
引用
收藏
页码:4070 / 4081
页数:12
相关论文
共 52 条
[1]   Translation elongation factor EF-Tu is a target for Stp, a serine-threonine phosphatase involved in virulence of Listeria monocytogenes [J].
Archambaud, C ;
Gouin, E ;
Pizarro-Cerda, J ;
Cossart, P ;
Dussurget, O .
MOLECULAR MICROBIOLOGY, 2005, 56 (02) :383-396
[2]   Control of Listeria superoxide dismutase by phosphorylation [J].
Archambaud, Cristel ;
Nahori, Marie-Anne ;
Pizarro-Cerda, Javier ;
Cossart, Pascale ;
Dussurget, Olivier .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (42) :31812-31822
[3]   New vector for efficient allelic replacement in naturally nontransformable, low-GC-content, gram-positive bacteria [J].
Arnaud, M ;
Chastanet, A ;
Débarbouillé, M .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2004, 70 (11) :6887-6891
[4]   The protein phosphatase 2C (PP2C) superfamily: Detection of bacterial homologues [J].
Bork, P ;
Brown, NP ;
Hegyi, H ;
Schultz, J .
PROTEIN SCIENCE, 1996, 5 (07) :1421-1425
[5]   Characterization of the Bacillus subtilis GTPase YloQ and its role in ribosome function [J].
Campbell, TL ;
Daigle, DM ;
Brown, ED .
BIOCHEMICAL JOURNAL, 2005, 389 :843-852
[6]   Regulation of Streptococcus pneumoniae clp genes and their role in competence development and stress survival [J].
Chastanet, A ;
Prudhomme, M ;
Claverys, JP ;
Msadek, T .
JOURNAL OF BACTERIOLOGY, 2001, 183 (24) :7295-7307
[7]   The GTPase, CpgA(YloQ), a putative translation factor, is implicated in morphogenesis in Bacillus subtilis [J].
Cladière, L ;
Hamze, K ;
Madec, E ;
Levdikov, VM ;
Wilkinson, AJ ;
Holland, IB ;
Séror, SJ .
MOLECULAR GENETICS AND GENOMICS, 2006, 275 (04) :409-420
[8]   Role of protein phosphorylation on serine/threonine and tyrosine in the virulence of bacterial pathogens [J].
Cozzone, AJ .
JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY, 2005, 9 (3-4) :198-213
[9]   Crystal structure of the protein serine/threonine phosphatase 2C at 2.0 angstrom resolution [J].
Das, AK ;
Helps, NR ;
Cohen, PTW ;
Barford, D .
EMBO JOURNAL, 1996, 15 (24) :6798-6809
[10]  
DONAT S, 2009, J BACTERIOL, DOI DOI 10.1128/JB0117-09