The proper ratio of GrpE to DnaK is important for protein quality control by the DnaK-DnaJ-GrpE chaperone system and for cell division

被引:44
作者
Sugimoto, Shinya [1 ,2 ]
Saruwatari, Kozue [1 ]
Higashi, Chihana [1 ]
Sonomoto, Kenji [1 ,2 ]
机构
[1] Kyushu Univ, Grad Sch, Fac Agr,Higashi Ku, Dept Biosci & Biotechnol,Div Microbial Sci & Tech, Fukuoka 8128581, Japan
[2] Kyushu Univ, Bio Architecture Ctr, Dept Funct Metab Design, Lab Funct Food Design,Hakozaki Ku, Fukuoka 8128581, Japan
来源
MICROBIOLOGY-SGM | 2008年 / 154卷
关键词
D O I
10.1099/mic.0.2008/017376-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A balance of the intracellular concentrations of molecular chaperones in response to environmental conditions is of considerable importance for cellular homeostasis. Here, the physiological consequences of overexpression of GrpE in wild-type Escherichia coli MC41 00 were examined. Overexpression of GrpE resulted in defects in cell division and growth, but overexpression of GrpE-G1 22D, which carries the G1 22D point mutation resulting in impaired interaction with DnaK, did not; this indicated that the effect of GrpE overexpression could be related to the DnaK chaperone function. Phase-contrast and fluorescence micrographs suggested that the N-terminal GFP-fused GrpE was colocalized with DnaK on the surface of inclusion bodies. An in vitro luciferase-refolding activity assay using purified DnaK, DnaJ and GrpE proteins demonstrated that high concentrations of GrpE significantly inhibited DnaK-mediated refolding. Furthermore, cell-free extracts from wild-type cells and GrpE-G1 221D-overexpressing cells significantly enhanced the refolding of luciferase. In the GrpE-overexpressing cells, abnormal localization of the cell-division protein FtsZ was observed by indirect immunofluorescence microscopy. In conclusion, the overexpression of GrpE caused a defect in the functionality of the DnaK chaperone system; this would result in filamentous morphology via abnormalities in the cell-division machinery.
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页码:1876 / 1885
页数:10
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