Molecular and General Genetics MGG

, Volume 199, Issue 3, pp 381–387 | Cite as

Suppressors of temperature-sensitive mutations in a ribosomal protein gene, rpsL (S12), of Escherichia coli K12

  • Hiroko Nashimoto
  • Akiko Miura
  • Haruo Saito
  • Hisao Uchida
Article

Summary

Temperature-sensitive (ts) mutations were isolated within a ribosomal protein gene (rpsL) of Escherichia coli K12. Mutations were mapped by complementation using various transducing phages and plasmids carrying the rpsL gene, having either a normal or a defective promoter for the rpsL operon. One of these mutations, ts118, resulted in a mutant S12 protein which behaved differently from the wild-type S12 on CM-cellulose column chromatography. Suppressors of these ts mutations were isolated and characterized; one was found to be a mutation of a nonribosomal protein gene which was closely linked to the RNAase III gene on the E. coli chromosome. This suppressor, which was recessive to its wild-type allele, was cloned into a transducing phage and mapped finely. A series of cold-sensitive mutations, affecting the assembly of ribosomes at 20°C, was isolated within the purL to nadB region of the E. coli chromosome and one group, named rbaA, mapped at the same locus as the suppressor mutation, showing close linkage to the RNAase III gene.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Adler J, Kaiser AD (1963) Mapping of the galactose genes of Escherichia coli by transduction with phage P1. Virology 19:117–126Google Scholar
  2. Bachmann BJ (1983) Linkage map of Escherichia coli K-12, Edition 7. Microbiol Rev 47:180–230Google Scholar
  3. Cowgill de Narvaez C, Schaup HW (1979) In vivo transcriptionally coupled assembly of Escherichia coli ribosomal subunits. J Mol Biol 134:1–22Google Scholar
  4. Dean D (1981) A plasmid cloning vector for the direct selection of strains carrying recombinant plasmids. Gene 15:99–102Google Scholar
  5. Dunn JJ, Studier FW (1973) T7 early RNAs and Escherihia coli ribosomal RNAs are cut from large precursor RNAs in vivo by ribonuclease III. Proc Natl Acad Sci USA 70:3296–3300Google Scholar
  6. Gegenheimer P, Apirion D (1981) Processing of procaryotic ribonucleic acid. Microbiol Rev 45:502–541Google Scholar
  7. Guthrie C, Nashimoto H, Nomura M (1969) Structure and function of E. coli ribosomes, VIII. Cold-sensitive mutants defective in ribosome assembly. Proc Natl Acad Sci USA 63:384–391Google Scholar
  8. Hartman PE, Roth JR (1973) Mechanisms of suppression. Adv Genet 17:1–105Google Scholar
  9. Hong JS, Ames BN (1971) Localized mutagenesis of any specific small region of the bacterial chromosome. Proc Natl Acad Sci USA 68:3158–3162Google Scholar
  10. Ishihama A, Shimamoto N, Aiba H, Kawakami K, Nashimoto H, Tsugawa A, Uchida H (1980) Temperature-sensitive mutations in the alpha subunit gene of Escherichia coli RNA polymerase. J Mol Biol 137:137–150Google Scholar
  11. Ito K, Wittekind M, Nomura M, Shiba K, Yura T, Miura A, Nashimoto H (1983) A temperature-sensitive mutant of E. coli exhibiting slow processing of exported proteins. Cell 32:789–797Google Scholar
  12. Kaltschmidt E, Wittmann HG (1970) Ribosomal proteins, VII. Two dimensional polyacrylamide gel electrophoresis for finger-printing of ribosomal proteins. Anal Biochem 36:401–412Google Scholar
  13. Kindler P, Keil TU, Hofschneider PH (1973) Isolation and characterization of an RNase III deficient mutant of Escherichia coli. Mol Gen Genet 126:53–69Google Scholar
  14. Leboy PS, Cox EC, Flaks JG (1964) The chromosomal site specifying a ribosomal protein in Escherichia coli. Proc Natl Acad Sci USA 52:1367–1374Google Scholar
  15. Lederberg J (1951) Streptomycin resistance: A genetically recessive mutation. J Bacteriol 61:549–550Google Scholar
  16. Lindahl L, Post L, Zengel J, Gilbert SF, Strycharz WA, Nomura M (1977) Mapping of ribosomal protein genes by in vitro protein synthesis using DNA fragments of λfus3 transducing phage DNA as templates. J Biol Chem 252:7365–7383Google Scholar
  17. Low B (1968) Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci USA 60:160–169Google Scholar
  18. Low KB (1972) Escherichia coli K-12 F-prime factors, old and new. Bacteriol Rev 36:587–607Google Scholar
  19. Murray NE, Murray K (1974) Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature 251:476–481Google Scholar
  20. Nashimoto H, Nomura M (1970) Structure and function of bacterial ribosomes, XI. Dependence of 50S ribosomal assembly on simultaneous assembly of 30S subunits. Proc Natl Acad Sci USA 67:1440–1447Google Scholar
  21. Nashimoto H, Uchida H (1975) Late steps in the assembly of 30S ribosomal proteins in vivo in a spectinomycin-resistant mutant of Escherichia coli. J Mol Biol 96:443–453Google Scholar
  22. Nashimoto H, Held W, Kaltschmidt E, Nomura M (1971) Structure and function of bacterial ribosomes XII. Accumulation of 21S particles by some cold-sensitive mutants of Escherichia coli. J Mol Biol 62:121–138Google Scholar
  23. Nikolaev N, Silengo L, Schlessinger D (1973) Synthesis of a large precursor RNA in a mutant of Escherichia coli. Proc Natl Acad Sci USA 70:3361–3365Google Scholar
  24. Nomura M (1970) Bacterial ribosomes. Bacteriol Rev 34:228–277Google Scholar
  25. Olins PO, Nomura M (1981) Regulation of the S10 ribosomal protein operon in E. coli: Nucleotide sequence at the start of the operon. Cell 26:205–211Google Scholar
  26. Parkinson JS, Huskey RJ (1971) Deletion mutants of bacteriophage lambda. J Mol Biol 56:369–384Google Scholar
  27. Pittard J, Wallace BJ (1966) Distribution and function of genes concerned with aromatic biosynthesis in Escherichia coli. J Bacteriol 91:1494–1508Google Scholar
  28. Post LE, Arfsten AE, Nomura M, Jaskunas SR (1978) Isolation and characterization of a promoter mutant in the str ribosomal protein operon in E. coli. Cell 15:231–236Google Scholar
  29. Saito H, Nakamura Y, Uchida H (1978) A transducing lambda phage carrying grpE, a bacterial gene necessary for lambda DNA replication, and two ribosomal protein genes, rpsP (S16) and rplS (L19). Mol Gen Genet 165:247–256Google Scholar
  30. Saito H, Uchida H (1978) Organization and expression of the dnaJ and dnaK genes of Escherichia coli K12. Mol Gen Genet 164:1–8Google Scholar
  31. Schrenk WJ, Weisberg RA (1975) A simple method for making new transducing lines of coliphage lambda. Mol Gen Genet 137:101–107Google Scholar
  32. Traub P, Nomura M (1969) Structure and function of E. coli ribosomes. VI. Mechanism of assembly of 30S ribosomal particles in vitro. J Mol Biol 40:391–413Google Scholar
  33. Tessman I (1968) Mutagenic treatment of double-and single-stranded DNA phage T4 and S13 with hydroxylamine. Virology 35:330–333Google Scholar
  34. Yochem J, Uchida H, Sunshine M, Saito H, Georgopoulos CP, Feiss M (1978) Genetic analysis of two genes, dnaJ and dnaK, necessary for Escherichia coli and bacteriophage lambda DNA replication. Mol Gen Genet 164:9–14Google Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Hiroko Nashimoto
    • 1
  • Akiko Miura
    • 1
  • Haruo Saito
    • 1
  • Hisao Uchida
    • 1
  1. 1.Institute of Medical ScienceUniversity of TokyoTokyoJapan

Personalised recommendations