Chaperonins as potential gene regulatory factors. In vitro interaction and solubilization of NifA, the nif transcriptional activator, with GroEL.
A previous study (Govezensky, D., Greener, T., and Zamir, A. (1991) J. Bacteriol. 20, 6339-6346) indicated that the chaperonin GroEL was required for maximal expression from nif promoters in Klebsiella pneumoniae and nif-transformed Escherichia coli. That this requirement stemmed from the ability of GroEL to properly fold NifA, the nif transcriptional activator, was first supported by co-immunoprecipitation of NifA in K. pneumoniae extracts with anti-GroEL antibodies. In the present in vitro study, NifA, partially purified from E. coli overexpressing the protein, was diluted from a 6 M urea solution into a refolding buffer in the presence or absence of GroEL. Dilution in the absence of GroEL caused the complete precipitation of NifA. When present in the dilution buffer, GroEL bound NifA and maintained it in a soluble state. GroEL was also found to bind NifA newly synthesized in an in vitro translation system. For both NifA preparations, cochaperonin GroES and ATP promoted release of NifA from GroEL. These results provide evidence for the association of NifA with GroEL and for the role of both GroEL and GroES in the solubilization and thereby folding of the nif transcriptional activator.
Govezensky
D
D
Department of Structural Biology, Weizmann Institute of Science, Rehovot, Israel.
Bochkareva
E S
ES
Zamir
A
A
Girshovich
A S
AS
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018834:Chaperonin 60; D018833:Chaperonins; D004926:Escherichia coli / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D007711:Klebsiella pneumoniae / Q000235:genetics; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D011506:Proteins / Q000502:physiology*; D012995:Solubility; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; C061338:NifA protein, Bacteria; D011506:Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D018833:Chaperonins