The Clp proteases of Bacillus subtilis are directly involved in degradation of misfolded proteins

被引:131
作者
Krüger, E [1 ]
Witt, E [1 ]
Ohlmeier, S [1 ]
Hanschke, R [1 ]
Hecker, M [1 ]
机构
[1] Ernst Moritz Arndt Univ Greifswald, Inst Mikrobiol & Mol Biol, D-17487 Greifswald, Germany
关键词
D O I
10.1128/JB.182.11.3259-3265.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The presence of the heat stress response-related ATPases ClpC and ClpX or the peptidase ClpP in the cell is crucial for tolerance of many forms of stress in Bacillus subtilis. Assays for detection of defects in protein degradation suggest that ClpC, ClpP, and ClpX participate directly in overall proteolysis of misfolded proteins. Turnover rates for abnormal puromycyl peptides are significantly decreased in clpC, clpP, and clpX mutant cells. Electron-dense aggregates, most likely due to the accumulation of misfolded proteins, were noticed in studies of ultrathin cryosections in clpC and clpP mutant cells even under nonstress conditions. In contrast, in the wild type or clpX mutants such aggregates could only be observed after heat shock. This phenomenon supports the assumption that clpC and clpP mutants are deficient in the ability to solubilize or degrade damaged and aggregated proteins, the accumulation of which is toxic for the cell. By using immunogold labeling with antibodies raised against ClpC, ClpP, and ClpX, the Clp proteins were localized in these aggregates,showing that the Clp proteins act at this level in vivo.
引用
收藏
页码:3259 / 3265
页数:7
相关论文
共 54 条
[1]   Crystal structure of heat shock locus V (HslV) from Escherichia coli [J].
Bochtler, M ;
Ditzel, L ;
Groll, M ;
Huber, R .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (12) :6070-6074
[2]   CONSTRUCTION AND CHARACTERIZATION OF NEW CLONING VEHICLES .2. MULTIPURPOSE CLONING SYSTEM [J].
BOLIVAR, F ;
RODRIGUEZ, RL ;
GREENE, PJ ;
BETLACH, MC ;
HEYNEKER, HL ;
BOYER, HW ;
CROSA, JH ;
FALKOW, S .
GENE, 1977, 2 (02) :95-113
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   PROTEOLYSIS AND MODULATION OF THE ACTIVITY OF THE CELL-DIVISION INHIBITOR SULA IN ESCHERICHIA-COLI LON MUTANTS [J].
CANCEILL, D ;
DERVYN, E ;
HUISMAN, O .
JOURNAL OF BACTERIOLOGY, 1990, 172 (12) :7297-7300
[5]   ClpP participates in the degradation of misfolded protein in Lactococcus lactis [J].
Frees, D ;
Ingmer, H .
MOLECULAR MICROBIOLOGY, 1999, 31 (01) :79-87
[6]   Sequence and transcriptional analysis of clpX, a class-III heat-shock gene of Bacillus subtilis [J].
Gerth, U ;
Wipat, A ;
Harwood, CR ;
Carter, N ;
Emmerson, PT ;
Hecker, M .
GENE, 1996, 181 (1-2) :77-83
[7]   Stress induction of the Bacillus subtilis clpP gene encoding a homologue of the proteolytic component of the Clp protease and the involvement of ClpP and ClpX in stress tolerance [J].
Gerth, U ;
Krüger, E ;
Derré, I ;
Msadek, T ;
Hecker, M .
MOLECULAR MICROBIOLOGY, 1998, 28 (04) :787-802
[9]   Regulatory subunits of energy-dependent proteases [J].
Gottesman, S ;
Maurizi, MR ;
Wickner, S .
CELL, 1997, 91 (04) :435-438
[10]   Protein quality control: Triage by chaperones and proteases [J].
Gottesman, S ;
Wickner, S ;
Maurizi, MR .
GENES & DEVELOPMENT, 1997, 11 (07) :815-823