The DnaK homologue of the marine Vibrio sp. strain S14 binds to the unprocessed form of a carbon starvation-specific periplasmic protein.
The Escherichia coli DnaK homologue in Vibrio sp. strain S14 was shown to possess chaperone function for translocation during carbon starvation. This was demonstrated by using the method of co-immunoprecipitation. DnaK co-precipitated with the carbon starvation-specific periplasmic space protein Csp5 three hours after the onset of carbon starvation. Pulse-chasing of the protein with radiolabelled methionine followed by the addition of an excess of unlabelled methionine demonstrated that the Csp5 protein was translocated across the inner membrane. Only the cytoplasmic unprocessed precursor form of Csp5 co-precipitated with DnaK. The non-covalent binding between the two proteins was found to be ATP-dependent, as the addition of ATP released the interaction between DnaK and the precursor form of Csp5, as was shown on silver-stained SDS-polyacrylamide gels and by Western blot analysis. We suggest that DnaK maintains the carbon starvation-inducible protein Csp5 in a translocation-competent form in the cytoplasm.
Holmquist
L
L
Department of General and Marine Microbiology, University of Göteborg, Sweden.
Nelson
D R
DR
Kjelleberg
S
S
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001692:Biological Transport; D002244:Carbon / Q000378:metabolism*; D018833:Chaperonins; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D017510:Protein Folding; D011506:Proteins / Q000378:metabolism*; D014733:Vibrio / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D008565:Membrane Proteins; D011506:Proteins; D002244:Carbon; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D018833:Chaperonins; C063842:dnaK protein, E coli