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. 1977 Oct;132(1):254–261. doi: 10.1128/jb.132.1.254-261.1977

Ribonucleic Acid Polymerase Mutant of Escherichia coli Defective in Flagella Formation

T Yamamori 1, K Ito 1, T Yura 1, T Suzuki 2, T Iino 2
PMCID: PMC221851  PMID: 199575

Abstract

Escherichia coli K-12 mutants that are resistant to bacteriophage χ, defective in motility, and unable to grow at high temperature (42°C) were isolated from among those selected for rifampin resistance at low temperature (30°C) after mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine. Genetic analysis of one such mutant indicated the presence of two mutations that probably affect the β subunit of ribonucleic acid (RNA) polymerase: one (rif) causing rifampin resistance and the other (Ts-74) conferring resistance to phage χ (and loss of motility) and temperature sensitivity for growth. Observations with an electron microscope revealed that the number of flagella per mutant cell was significantly reduced, suggesting that the Ts-74 mutation somehow affected flagella formation at the permissive temperature. When a mutant culture was transferred from 30 to 42°C, deoxyribonucleic acid synthesis accelerated normally, but RNA or protein synthesis was enhanced relatively little. The rate of synthesis of β and β′ subunits of RNA polymerase was low even at 30°C and was further reduced at 42°C, in contrast to the parental wild-type strain. Expression of the lactose and other sugar fermentation operons, as well as lysogenization with phage λ, occurred normally at 30°C, suggesting that the mutation does not cause general shut-off of gene expression regulated by cyclic adenosine 3′,5′-monophosphate.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Burgess R. R. Separation and characterization of the subunits of ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6168–6176. [PubMed] [Google Scholar]
  4. Errington L., Glass R. E., Hayward R. S., Scaife J. G. Structure and orientation of an RNA polymerase operon in Escherichia coli. Nature. 1974 Jun 7;249(457):519–522. doi: 10.1038/249519a0. [DOI] [PubMed] [Google Scholar]
  5. Georgopoulos C. P. Bacterial mutants in which the gene N function of bacteriophage lambda is blocked have an altered RNA polymerase. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2977–2981. doi: 10.1073/pnas.68.12.2977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gudas L. J., James R., Paradee A. B. Evidence of the involvement of an outer membrane protein in DNA initiation. J Biol Chem. 1976 Jun 10;251(11):3470–3479. [PubMed] [Google Scholar]
  7. Heil A., Zillig W. Reconstitution of bacterial DNA-dependent RNA-polymerase from isolated subunits as a tool for the elucidation of the role of the subunits in transcription. FEBS Lett. 1970 Dec;11(3):165–168. doi: 10.1016/0014-5793(70)80519-1. [DOI] [PubMed] [Google Scholar]
  8. Hong J. S., Smith G. R., Ames B. N. Adenosine 3':5'-cyclic monophosphate concentration in the bacterial host regulates the viral decision between lysogeny and lysis. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2258–2262. doi: 10.1073/pnas.68.9.2258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Howard-Flanders P., Boyce R. P., Theriot L. Three loci in Escherichia coli K-12 that control the excision of pyrimidine dimers and certain other mutagen products from DNA. Genetics. 1966 Jun;53(6):1119–1136. doi: 10.1093/genetics/53.6.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Iino T. Genetics and chemistry of bacterial flagella. Bacteriol Rev. 1969 Dec;33(4):454–475. doi: 10.1128/br.33.4.454-475.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ikeda H., Tomizawa J. I. Transducing fragments in generalized transduction by phage P1. I. Molecular origin of the fragments. J Mol Biol. 1965 Nov;14(1):85–109. doi: 10.1016/s0022-2836(65)80232-7. [DOI] [PubMed] [Google Scholar]
  12. Ikeuchi T., Yura T., Yamagishi H. Genetic and physical studies of lambda transducing bacteriophage carrying the beta subunit gene of the Escherichia coli ribonucleic acid polymerase. J Bacteriol. 1975 Jun;122(3):1247–1256. doi: 10.1128/jb.122.3.1247-1256.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Iwakura Y., Ishihama A., Yura T. RNA polymerase mutants of Escherichia coli. Streptolydigin resistance and its relation to rifampicin resistance. Mol Gen Genet. 1973 Mar 1;121(2):181–196. doi: 10.1007/BF00277531. [DOI] [PubMed] [Google Scholar]
  14. Iwakura Y., Ito K., Ishihama A. Biosynthesis of RNA polymerase in Escherichia coli. I. Control of RNA polymerase content at various growth rates. Mol Gen Genet. 1974;133(1):1–23. doi: 10.1007/BF00268673. [DOI] [PubMed] [Google Scholar]
  15. Kirschbaum J. B., Claeys I. V., Nasi S., Molholt B., Miller J. H. Temperature-sensitive RNA polymerase mutants with altered subunit synthesis and degradation. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2375–2379. doi: 10.1073/pnas.72.6.2375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kirschbaum J. B., Scaife J. Evidence for a lambda transducing phage carrying the genes for the beta and beta' subunits of Escherichia coli RNA polymerase. Mol Gen Genet. 1974;132(3):193–201. doi: 10.1007/BF00269392. [DOI] [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. MEYNELL E. W. A phage, phi chi, which attacks motile bacteria. J Gen Microbiol. 1961 Jun;25:253–290. doi: 10.1099/00221287-25-2-253. [DOI] [PubMed] [Google Scholar]
  19. Morishita T., Yura T. Altered nutritional requirements associated with mutations affecting the structures of ribonucleic acid polymerase in Lactobacillus casei. J Bacteriol. 1976 Feb;125(2):416–422. doi: 10.1128/jb.125.2.416-422.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nakamura Y., Yura T. Evidence for a positive regulation of RNA polymerase synthesis in Escherichia coli. J Mol Biol. 1975 Oct 5;97(4):621–642. doi: 10.1016/s0022-2836(75)80063-5. [DOI] [PubMed] [Google Scholar]
  21. Nakamura Y., Yura T. Hyperproduction of the sigma subunit of RNA polymerase in a mutant of Escherichia coli. Mol Gen Genet. 1975 Nov 24;141(2):97–111. doi: 10.1007/BF00267677. [DOI] [PubMed] [Google Scholar]
  22. Oeschger M. P., Berlyn M. K. Regulation of RNA polymerase synthesis in Escherichia coli: a mutant unable to synthesize the enzyme at 43 degrees. Proc Natl Acad Sci U S A. 1975 Mar;72(3):911–915. doi: 10.1073/pnas.72.3.911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Oki M. Correlation between metabolism of phosphatidylglycerol and membrane synthesis in Escherichia coli. J Mol Biol. 1972 Jul 21;68(2):249–264. doi: 10.1016/0022-2836(72)90212-4. [DOI] [PubMed] [Google Scholar]
  24. Ryter A., Shuman H., Schwartz M. Intergration of the receptor for bacteriophage lambda in the outer membrane of Escherichia coli: coupling with cell division. J Bacteriol. 1975 Apr;122(1):295–301. doi: 10.1128/jb.122.1.295-301.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Schade S., Adler J. Purification and chemistry of bacteriophage chi. J Virol. 1967 Jun;1(3):591–598. doi: 10.1128/jvi.1.3.591-598.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shen B. H., Boos W. Regulation of the -methylgalactoside transport system and the galatose-binding protein by the cell cycle of Escherichia coli. Proc Natl Acad Sci U S A. 1973 May;70(5):1481–1485. doi: 10.1073/pnas.70.5.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Silerman M., Matsumura P., Draper R., Edwards S., Simon M. I. Expression of flagellar genes carried by bacteriophage lambda. Nature. 1976 May 20;261(5557):248–250. doi: 10.1038/261248a0. [DOI] [PubMed] [Google Scholar]
  28. Silverman M., Simon M. Characterization of Escherichia coli flagellar mutants that are insensitive to catabolite repression. J Bacteriol. 1974 Dec;120(3):1196–1203. doi: 10.1128/jb.120.3.1196-1203.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sonenshein A. L., Losick R. RNA polymerase mutants blocked in sporulation. Nature. 1970 Aug 29;227(5261):906–909. doi: 10.1038/227906a0. [DOI] [PubMed] [Google Scholar]
  30. Suzuki H., Iino T. Absence of messenger ribonucleic acid specific for flagellin in non-flagellate mutants of Salmonella. J Mol Biol. 1975 Jul 15;95(4):549–556. doi: 10.1016/0022-2836(75)90316-2. [DOI] [PubMed] [Google Scholar]
  31. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  32. Yokota T., Gots J. S. Requirement of adenosine 3', 5'-cyclic phosphate for flagella formation in Escherichia coli and Salmonella typhimurium. J Bacteriol. 1970 Aug;103(2):513–516. doi: 10.1128/jb.103.2.513-516.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]

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