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

Shinya Sugimoto, Kozue Saruwatari, Chihana Higashi, Kenji Sonomoto

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

A balance of the intracellular concentrations of molecular chaperenes 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 MC4100 were examined. Overexpression of GrpE resulted in defects in cell division and growth, but overexpression of GrpE-G1 22D, 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 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-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.

Original languageEnglish
Pages (from-to)1876-1885
Number of pages10
JournalMicrobiology
Volume154
Issue number7
DOIs
Publication statusPublished - Sep 11 2008

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Quality Control
Cell Division
Luciferases
HSP40 Heat-Shock Proteins
Proteins
Inclusion Bodies
Indirect Fluorescent Antibody Technique
Cell Extracts
Fluorescence Microscopy
Point Mutation
Homeostasis
Fluorescence
Escherichia coli
Growth
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Microbiology

Cite this

The proper ratio of GrpE to Dnak is important for protein quality control by the Dnak-DnaJ-GrpE chaperone system and for cell division. / Sugimoto, Shinya; Saruwatari, Kozue; Higashi, Chihana; Sonomoto, Kenji.

In: Microbiology, Vol. 154, No. 7, 11.09.2008, p. 1876-1885.

Research output: Contribution to journalArticle

Sugimoto, Shinya ; Saruwatari, Kozue ; Higashi, Chihana ; Sonomoto, Kenji. / The proper ratio of GrpE to Dnak is important for protein quality control by the Dnak-DnaJ-GrpE chaperone system and for cell division. In: Microbiology. 2008 ; Vol. 154, No. 7. pp. 1876-1885.
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