Summary
A suppressor mutation of a temperature-sensitive mutant of ribosomal protein L24 (rplX19) was mapped close to the lon gene by genetic analysis and was shown to affect protease LA. The degradation and the synthesis rates of individual ribosomal proteins were determined. Proteins L24, L14, L15 and L27 were found to be degraded faster in the original rplX19 mutant than in the rplX19 mutant containing the suppressor mutation. Other ribosomal proteins were either weakly or not at all degraded in both mutants. Temperature-sensitive growth was also suppressed by the overproduction of mutant protein L24 from a plasmid. Our results suggest that (1) either free ribosomal proteins or proteins bound to abortive assembly precursors are highly susceptible to the lon gene product and (2) the mutationally altered protein L24 can still function at the nonpermissive growth temperature of the mutant, if it is present in sufficient amounts.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amann E, Brosius J, Ptashne M (1983) Vectors bearing a hybrid trp-lac promoter useful for regulated expression of cloned genes in Escherichia coli. Gene 25:167–178
Branlant C, Sriwidada J, Krol A, Ebel JP (1977) RNA sequences in ribonucleoprotein fragments of the complex formed from ribosomal 23S RNA and ribosomal protein L24 of Escherichia coli. Eur J Biochem 74:155–170
Brosius J, Dull TJ, Sleeter DD, Noller HF (1981) Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli J Mol Biol 148:107–127
Cabezon T, Herzog A, Petre J, Yaguchi M, Bollen A (1977) Ribosomal assembly deficiency in an Escherichia coli thermosensitive mutant having an altered L24 ribosomal protein. J Mol Biol 116:361–374
Chung CH, Goldberg AL (1981) The product of the lon gene (capR) in E. coli is the ATP-dependent protease LA. Proc Natl Acad Sci USA 78:4931–4935
deBoer HA, Comstock LJ, Vasser M (1983) The tac promoter: A functional hybrid derived from trp and lac promoters. Proc Natl Acad Sci USA 80:21–25
Geyl D, Böck A, Isono K (1981) An improved method for twodimensional gel-gelectrophoresis analysis of mutationally altered ribosomal proteins of Escherichia coli. Mol Gen Genet 181:309–312
Goff SA, Casson LP, Goldberg AL (1984) Heat shock regulatory gene htpR influences rates of protein degradation and expression of the lon gene in Escherichia coli. Proc Natl Acad Sci USA 81:6647–6651
Goldberg AL (1972) Degradation of abnormal proteins in E. coli. Proc Natl Acad Sci USA 69:422–426
Gottesmann S, Zipser D (1978) Deg phenotype of Escherichia coli lon mutants. J Bacteriol 133:844–851
Grossmann AD, Burgess RR, Walter W, Gross CA (1983) Mutations in the lon gene of E. coli K 12 phenotypically suppress a mutation in the sigma subunit of RNA polymerase. Cell 32:151–159
Herold M, Nierhaus KH (1987) Incorporation of six additional proteins, to complete the assembly map of the 50 S subunit from Escherichia coli ribosomes. J Biol Chem in press
Howard-Flanders P, Simson E, Theriot L (1964) A locus that controls filament formation and sensitivity to radiation in Escherichia coli K 12. Genetics 49:237–246
Hua S, Markovitz A (1972) Multiple regulatory gene control of the galactose operon in Escherichia coli K 12. J Bacteriol 110:1089–1099
Kirby EP, Ruff WL, Goldthwait DA (1972) Cell division and prophage induction in E. coli: effects of pantoyl lactone and various furan derivatives. J Bacteriol 111:447–453
Lemaux PG, Herendeen SL, Bloch PL, Neidhardt FC (1978) Transient rates of synthesis of individual polypeptides in E. coli following temperature shifts. Cell 13:427–434
Lindahl L, Zengel JM (1979) Operon specific regulation of ribosomal protein synthesis in Escherichia coli. Proc Natl Acad Sci USA 76:6542–6546
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, New York
Neidhardt FC, Vaughn V, Phillips TA, Bloch PL (1983) Geneprotein index in Escherichia coli. Microbiol Rev 47:231–284
Nishi K, Schnier J (1986) A temperature-sensitive mutant in the rplX gene for ribosomal protein L24 and its suppression by spontaneous mutations in a 23 S rRNA gene of Escherichia coli. EMBO J 5:1373–1376
Nishi K, Dabbs ER, Schnier J (1985) DNA sequence and complementation analysis of a mutation in the rplX gene from Escherichia coli leading to loss of ribosomal protein L24. J Bacteriol 163:890–894
Nowotny V, Nierhaus KH (1982) Initiator proteins for the assembly of the 50 S subunit from Escherichia coli ribosomes. Proc Natl Acad Sci USA 79:7238–7242
O'Farrell PH (1975) High resolution two-dimensional eletrophoresis of proteins. J Biol Chem 250:4007–4021
Phillips TA, VanBogelen RA, Neidhardt FC (1984) The lon (CapR) gene product of Escherichia coli is a heat-shock protein. J Bacteriol 159:283–287
Sloof P, Hunter JB, Garrett RA, Branlant C (1978) RNA-RNA interactions in the binding site of protein L24 on 23S ribosomal RNA of Escherichia coli: 1. Evidence for their occurrence between widely spread separated sequence regions. Nucleic Acids Res 5:3503–3513
Takabe Y, Miura A, Bedwell DM, Tam M, Nomura M (1985) Increased expression of ribosomal genes during inhibition of ribosomal assembly in Escherichia coli. J Mol Biol 184:23–30
Zengel JM, Lindahl L (1985) Transcriptional control of the S10 ribosomal protein operon of Escherichia coli after a shift to higher temperature. J Bacteriol 163:140–147
Author information
Authors and Affiliations
Additional information
Communicated by J.W. Lengeler
Rights and permissions
About this article
Cite this article
Nishi, K., Schnier, J. The phenotypic suppression of a mutation in the gene rplX for ribosomal protein L24 by mutations affecting the lon gene product for protease LA in Escherichia coli K12. Molec. Gen. Genet. 212, 177–181 (1988). https://doi.org/10.1007/BF00322462
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00322462