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1887

Microbiology Society logo Microbiology

Volume 154, Issue 7

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

Abstract

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 MC4100 were examined. Overexpression of GrpE resulted in defects in cell division and growth, but overexpression of GrpE-G122D, which carries the G122D 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 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-G122D-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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Keyword(s): CBB, Coomassie brilliant blue and DAPI, 4,6-diamidino-2-phenylindole
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2008-07-01
2025-01-16
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/content/journal/micro/10.1099/mic.0.2008/017376-0
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The proper ratio of GrpE to DnaK is important for protein quality control by the DnaK–DnaJ–GrpE chaperone system and for cell division
Microbiology 154, 1876 (2008); https://doi.org/10.1099/mic.0.2008/017376-0
/content/journal/micro/10.1099/mic.0.2008/017376-0
/content/journal/micro/10.1099/mic.0.2008/017376-0
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