Two classes of extragenic suppressor mutations identify functionally distinct regions of the GroEL chaperone of Escherichia coli.
The GroES and GroEL proteins of Escherichia coli function together as the GroE molecular chaperone machine to (i) prevent denaturation and aggregation and (ii) assist the folding and oligomerization of other proteins without being part of the final structure. Previous genetic and biochemical analyses have determined that this activity requires interactions of the GroES 7-mer with the GroEL 14-mer. Recently, we have identified a region of the GroES protein that interacts with the GroEL protein. To identify those residues of the GroEL protein that interact with GroES, we have exploited the thermosensitive phenotype of strains bearing mutations at one or the other of two GroEL-interacting residues of GroES. We have isolated, cloned, and sequenced six suppressor mutations in groEL, three independent isolates for each groES mutant. Changes of only three different amino acid substitutions in GroEL protein were found among these six groEL suppressor mutations. On the basis of a number of in vivo analyses of the chaperone activity of various combinations of groES mutant alleles and groEL suppressor alleles, we propose that an amino-proximal region of the GroEL protein which includes amino acid residues 174 and 190 interacts with GroES and that a carboxyl-proximal region which includes residue 375 interacts with substrate proteins.
Department of Cellular, Viral, and Molecular Biology, School of Medicine, University of Utah, Salt Lake City 84132.
J
J
Zeilstra-Ryalls
O
O
Fayet
C
C
Georgopoulos
D001426:Bacterial Proteins / Q000378:metabolism; D010582:Bacteriophage lambda / Q000378:metabolism; D018835:Chaperonin 10 / Q000378:metabolism*; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000821:virology; D011485:Protein Binding; D017510:Protein Folding; D017422:Sequence Analysis, DNA; D013329:Structure-Activity Relationship; D013489:Suppression, Genetic; D014764:Viral Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D004268:DNA-Binding Proteins; C066312:DnaA protein, Bacteria; D014764:Viral Proteins