Hsp 33 Controls Elongation Factor-Tu Stability and allows Escherichia coli growth in the absence of the major DnaK and Trigger Factor Chaperones

N. Bruel, M.P. Castanié-Cornet, A.M. Cirinesi, G.M. Koningstein, C. Georgopoulos, J. Luirink, P. Genevaux

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Intracellular de novo protein folding is assisted by cellular networks of molecular chaperones. In Escherichia coli, cooperation between the chaperones trigger factor (TF) and DnaK is central to this process. Accordingly, the simultaneous deletion of both chaperone-encoding genes leads to severe growth and protein folding defects. Herein, we took advantage of such defective phenotypes to further elucidate the interactions of chaperone networks in vivo. We show that disruption of the TF/DnaK chaperone pathway is efficiently rescued by overexpression of the redox-regulated chaperone Hsp33. Consistent with this observation, the deletion of hslO, the Hsp33 structural gene, is no longer tolerated in the absence of the TF/DnaK pathway. However, in contrast with other chaperones like GroEL or SecB, suppression by Hsp33 was not attributed to its potential overlapping general chaperone function(s). Instead, we show that overexpressed Hsp33 specifically binds to elongation factor-Tu (EF-Tu) and targets it for degradation by the protease Lon. This synergistic action of Hsp33 and Lon was responsible for the rescue of bacterial growth in the absence of TF and DnaK, by presumably restoring the coupling between translation and the downstream folding capacity of the cell. In support of this hypothesis, we show that overexpression of the stress-responsive toxin HipA, which inhibits EF-Tu, also rescues bacterial growth and protein folding in the absence of TF and DnaK. The relevance for such a convergence of networks of chaperones and proteases acting directly on EF-Tu to modulate the intracellular rate of protein synthesis in response to protein aggregation is discussed. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
Original languageEnglish
Pages (from-to)44435-44446
JournalJournal of Biological Chemistry
Volume287
DOIs
Publication statusPublished - 2012

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Peptide Elongation Factor Tu
Protein folding
Protein Folding
Escherichia coli
Protease La
Growth
Molecular Chaperones
Gene encoding
Bacterial Proteins
Genes
Oxidation-Reduction
Proteins
Peptide Hydrolases
Agglomeration
Phenotype
Degradation
Defects

Cite this

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title = "Hsp 33 Controls Elongation Factor-Tu Stability and allows Escherichia coli growth in the absence of the major DnaK and Trigger Factor Chaperones",
abstract = "Intracellular de novo protein folding is assisted by cellular networks of molecular chaperones. In Escherichia coli, cooperation between the chaperones trigger factor (TF) and DnaK is central to this process. Accordingly, the simultaneous deletion of both chaperone-encoding genes leads to severe growth and protein folding defects. Herein, we took advantage of such defective phenotypes to further elucidate the interactions of chaperone networks in vivo. We show that disruption of the TF/DnaK chaperone pathway is efficiently rescued by overexpression of the redox-regulated chaperone Hsp33. Consistent with this observation, the deletion of hslO, the Hsp33 structural gene, is no longer tolerated in the absence of the TF/DnaK pathway. However, in contrast with other chaperones like GroEL or SecB, suppression by Hsp33 was not attributed to its potential overlapping general chaperone function(s). Instead, we show that overexpressed Hsp33 specifically binds to elongation factor-Tu (EF-Tu) and targets it for degradation by the protease Lon. This synergistic action of Hsp33 and Lon was responsible for the rescue of bacterial growth in the absence of TF and DnaK, by presumably restoring the coupling between translation and the downstream folding capacity of the cell. In support of this hypothesis, we show that overexpression of the stress-responsive toxin HipA, which inhibits EF-Tu, also rescues bacterial growth and protein folding in the absence of TF and DnaK. The relevance for such a convergence of networks of chaperones and proteases acting directly on EF-Tu to modulate the intracellular rate of protein synthesis in response to protein aggregation is discussed. {\circledC} 2012 by The American Society for Biochemistry and Molecular Biology, Inc.",
author = "N. Bruel and M.P. Castani{\'e}-Cornet and A.M. Cirinesi and G.M. Koningstein and C. Georgopoulos and J. Luirink and P. Genevaux",
year = "2012",
doi = "10.1074/jbc.M112.418525",
language = "English",
volume = "287",
pages = "44435--44446",
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Hsp 33 Controls Elongation Factor-Tu Stability and allows Escherichia coli growth in the absence of the major DnaK and Trigger Factor Chaperones. / Bruel, N.; Castanié-Cornet, M.P.; Cirinesi, A.M.; Koningstein, G.M.; Georgopoulos, C.; Luirink, J.; Genevaux, P.

In: Journal of Biological Chemistry, Vol. 287, 2012, p. 44435-44446.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Hsp 33 Controls Elongation Factor-Tu Stability and allows Escherichia coli growth in the absence of the major DnaK and Trigger Factor Chaperones

AU - Bruel, N.

AU - Castanié-Cornet, M.P.

AU - Cirinesi, A.M.

AU - Koningstein, G.M.

AU - Georgopoulos, C.

AU - Luirink, J.

AU - Genevaux, P.

PY - 2012

Y1 - 2012

N2 - Intracellular de novo protein folding is assisted by cellular networks of molecular chaperones. In Escherichia coli, cooperation between the chaperones trigger factor (TF) and DnaK is central to this process. Accordingly, the simultaneous deletion of both chaperone-encoding genes leads to severe growth and protein folding defects. Herein, we took advantage of such defective phenotypes to further elucidate the interactions of chaperone networks in vivo. We show that disruption of the TF/DnaK chaperone pathway is efficiently rescued by overexpression of the redox-regulated chaperone Hsp33. Consistent with this observation, the deletion of hslO, the Hsp33 structural gene, is no longer tolerated in the absence of the TF/DnaK pathway. However, in contrast with other chaperones like GroEL or SecB, suppression by Hsp33 was not attributed to its potential overlapping general chaperone function(s). Instead, we show that overexpressed Hsp33 specifically binds to elongation factor-Tu (EF-Tu) and targets it for degradation by the protease Lon. This synergistic action of Hsp33 and Lon was responsible for the rescue of bacterial growth in the absence of TF and DnaK, by presumably restoring the coupling between translation and the downstream folding capacity of the cell. In support of this hypothesis, we show that overexpression of the stress-responsive toxin HipA, which inhibits EF-Tu, also rescues bacterial growth and protein folding in the absence of TF and DnaK. The relevance for such a convergence of networks of chaperones and proteases acting directly on EF-Tu to modulate the intracellular rate of protein synthesis in response to protein aggregation is discussed. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

AB - Intracellular de novo protein folding is assisted by cellular networks of molecular chaperones. In Escherichia coli, cooperation between the chaperones trigger factor (TF) and DnaK is central to this process. Accordingly, the simultaneous deletion of both chaperone-encoding genes leads to severe growth and protein folding defects. Herein, we took advantage of such defective phenotypes to further elucidate the interactions of chaperone networks in vivo. We show that disruption of the TF/DnaK chaperone pathway is efficiently rescued by overexpression of the redox-regulated chaperone Hsp33. Consistent with this observation, the deletion of hslO, the Hsp33 structural gene, is no longer tolerated in the absence of the TF/DnaK pathway. However, in contrast with other chaperones like GroEL or SecB, suppression by Hsp33 was not attributed to its potential overlapping general chaperone function(s). Instead, we show that overexpressed Hsp33 specifically binds to elongation factor-Tu (EF-Tu) and targets it for degradation by the protease Lon. This synergistic action of Hsp33 and Lon was responsible for the rescue of bacterial growth in the absence of TF and DnaK, by presumably restoring the coupling between translation and the downstream folding capacity of the cell. In support of this hypothesis, we show that overexpression of the stress-responsive toxin HipA, which inhibits EF-Tu, also rescues bacterial growth and protein folding in the absence of TF and DnaK. The relevance for such a convergence of networks of chaperones and proteases acting directly on EF-Tu to modulate the intracellular rate of protein synthesis in response to protein aggregation is discussed. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.

U2 - 10.1074/jbc.M112.418525

DO - 10.1074/jbc.M112.418525

M3 - Article

VL - 287

SP - 44435

EP - 44446

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

ER -