Trigger factor and DnaK cooperate in folding of newly synthesized proteins.
The role of molecular chaperones in assisting the folding of newly synthesized proteins in the cytosol is poorly understood. In Escherichia coli, GroEL assists folding of only a minority of proteins and the Hsp70 homologue DnaK is not essential for protein folding or cell viability at intermediate growth temperatures. The major protein associated with nascent polypeptides is ribosome-bound trigger factor, which displays chaperone and prolyl isomerase activities in vitro. Here we show that delta tig::kan mutants lacking trigger factor have no defects in growth or protein folding. However, combined delta tig::kan and delta dnaK mutations cause synthetic lethality. Depletion of DnaK in the delta tig::kan mutant results in massive aggregation of cytosolic proteins. In delta tig::kan cells, an increased amount of newly synthesized proteins associated transiently with DnaK. These findings show in vivo activity for a ribosome-associated chaperone, trigger factor, in general protein folding, and functional cooperation of this protein with a cytosolic Hsp70. Trigger factor and DnaK cooperate to promote proper folding of a variety of E. coli proteins, but neither is essential for folding and viability at intermediate growth temperatures.
Deuerling
E
E
Institut für Biochemie und Molekularbiologie, Freiburg, Germany.
Schulze-Specking
A
A
Tomoyasu
T
T
Mogk
A
A
Bukau
B
B
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism / Q000502:physiology*; D018834:Chaperonin 60 / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins / Q000502:physiology*; D007612:Kanamycin / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D019696:Peptidylprolyl Isomerase / Q000502:physiology*; D017510:Protein Folding
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D007612:Kanamycin; D008156:Luciferases; C063842:dnaK protein, E coli; D019696:Peptidylprolyl Isomerase