Previous Article | Next Article 
J Bacteriol. 1994 November; 176(22): 6980-6985
A carboxy-terminal deletion impairs the assembly of GroEL and confers a pleiotropic phenotype in Escherichia coli K-12.
B P Burnett,
A L Horwich and
K B Low
Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, Connecticut 06510.
ABSTRACT
A series of COOH-terminal deletions of the chaperonin GroEL have been examined for effects in vivo at haploid copy number on the essential requirement of GroEL for cell growth. Strains with a deletion of up to 27 COOH-terminal amino acids were viable, but not viable strain could be isolated with a deletion of 28 or more codons. When substitutions were placed in the COOH-terminal amino acid Val-521 of the 27-amino-acid-deleted (delta 27) mutant, we found variable effect--Trp and Glu led to inviability, whereas Arg and Gly were viable but slow growing. The effects of the Arg substitution plus deletion (V521R delta) were examined in more detail. Whereas the delta 27 mutant with the wild-type residue Val-521 grew as well as a strain with wild-type GroEL, the V521R delta mutant strain (groEL202) exhibited a broad range of phenotypic defects. These include slow growth; filamentous morphology; a defect in plating lambda; absence of activity of expressed human ornithine transcarbamylase, as seen in other GroEL mutants; and several newly observed defects, such as absence of motility, sensitivity to UV light and mitomycin, a defect in one mode of specialized transduction, and inability to grow on rhamnose. Sucrose gradient analysis of extracts from the V521R delta cells showed a substantially reduced level of GroEL sedimenting at the normal 20S position of the assembled tetradecamer and a relatively large amount of more lightly sedimenting subunits. This indicates that the substitution-deletion mutation interferes with oligomeric assembly of GroEL into its functional form. This is discussed in light of the recently determined crystal structure of GroEL.
J Bacteriol. 1994 November; 176(22): 6980-6985
This article has been cited by other articles:
-
Suzuki, M., Ueno, T., Iizuka, R., Miura, T., Zako, T., Akahori, R., Miyake, T., Shimamoto, N., Aoki, M., Tanii, T., Ohdomari, I., Funatsu, T.
(2008). Effect of the C-terminal Truncation on the Functional Cycle of Chaperonin GroEL: IMPLICATION THAT THE C-TERMINAL REGION FACILITATES THE TRANSITION FROM THE FOLDING-ARRESTED TO THE FOLDING-COMPETENT STATE. J. Biol. Chem.
283: 23931-23939
[Abstract]
[Full Text]
-
Machida, K., Kono-Okada, A., Hongo, K., Mizobata, T., Kawata, Y.
(2008). Hydrophilic Residues 526KNDAAD531 in the Flexible C-terminal Region of the Chaperonin GroEL Are Critical for Substrate Protein Folding within the Central Cavity. J. Biol. Chem.
283: 6886-6896
[Abstract]
[Full Text]
-
Piao, Z., Sze, C. C., Barysheva, O., Iida, K.-i., Yoshida, S.-i.
(2006). Temperature-Regulated Formation of Mycelial Mat-Like Biofilms by Legionella pneumophila. Appl. Environ. Microbiol.
72: 1613-1622
[Abstract]
[Full Text]
-
Park, S. G., Jung, G.
(2001). Human Hepatitis B Virus Polymerase Interacts with the Molecular Chaperonin Hsp60. J. Virol.
75: 6962-6968
[Abstract]
[Full Text]
-
Hogenhout, S. A., van der Wilk, F., Verbeek, M., Goldbach, R. W., van den Heuvel, J. F. J. M.
(2000). Identifying the Determinants in the Equatorial Domain of Buchnera GroEL Implicated in Binding Potato Leafroll Virus. J. Virol.
74: 4541-4548
[Abstract]
[Full Text]
-
Berlyn, M. K. B.
(1998). Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Microbiol. Mol. Biol. Rev.
62: 814-984
[Abstract]
[Full Text]
-
Baldini, R. L., Avedissian, M., Gomes, S. L.
(1998). The CIRCE Element and Its Putative Repressor Control Cell Cycle Expression of the Caulobacter crescentus groESL Operon. J. Bacteriol.
180: 1632-1641
[Abstract]
[Full Text]
-
Hogenhout, S. A., van der Wilk, F., Verbeek, M., Goldbach, R. W., van den Heuvel, J. F. J. M.
(1998). Potato Leafroll Virus Binds to the Equatorial Domain of the Aphid Endosymbiotic GroEL Homolog. J. Virol.
72: 358-365
[Abstract]
[Full Text]
-
White, Z. W., Fisher, K. E., Eisenstein, E.
(1995). A Monomeric Variant of GroEL Binds Nucleotides but Is Inactive as a Molecular Chaperone. J. Biol. Chem.
270: 20404-20409
[Abstract]
[Full Text]
-
Galan, A., Sot, B., Llorca, O., Carrascosa, J. L., Valpuesta, J. M., Muga, A.
(2001). Excluded Volume Effects on the Refolding and Assembly of an Oligomeric Protein. GroEL, A CASE STUDY. J. Biol. Chem.
276: 957-964
[Abstract]
[Full Text]
Copyright © 1994 by the American Society for Microbiology. All rights reserved.