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. 1976 Jul;127(1):32–39. doi: 10.1128/jb.127.1.32-39.1976

Wild-type and mutant in vitro products of an operon for ribonucleic acid polymerase subunits.

S Austin
PMCID: PMC233029  PMID: 776928

Abstract

An in vitro protein-synthesizing system can synthesize two ribonucleic acid (RNA) polymerase subunits of Escherichia coli, beta and beta', when a transducing phage deoxyribonucleic acid (DNA) template containing the rpoB region of the bacterial chromosome is added. Recombinant rpoB transducing phages were isolated that carry "nonsense" mutations of the class rpo-rifampin zero amber (formally referred to as rifoam). DNA was extracted from two of these phages. These DNAs are unable to direct the synthesis of beta subunits, whereas beta' synthesis is unaffected. Both mutations can be efficiently suppressed in vitro by the addition of suppressor transfer RNA. One of the mutations (rpoB115) produces a detectable nonsense (or restart) fragment of the beta protein in the absence of suppression. It is concluded that rpoB115 is an amber mutation within the structural gene for the beta subunit of RNA polymerase.

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Selected References

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

  1. Austin S. J., Tittawella I. P., Hayward R. S., Scaife J. G. Amber mutations of Escherichia coli RNA polymerase. Nat New Biol. 1971 Aug 4;232(31):133–136. doi: 10.1038/newbio232133a0. [DOI] [PubMed] [Google Scholar]
  2. Austin S. Coordinate and differential in vitro syntheses of two RNA polymerase subunits. Nature. 1974 Dec 13;252(5484):596–597. doi: 10.1038/252596a0. [DOI] [PubMed] [Google Scholar]
  3. Austin S., McGeoch D. The synthesis in vitro of RNA polymerase subunits of Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2420–2423. doi: 10.1073/pnas.70.8.2420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Austin S., Scaife J. A new method for selecting RNA polymerase mutants. J Mol Biol. 1970 Apr 14;49(1):263–267. doi: 10.1016/0022-2836(70)90394-3. [DOI] [PubMed] [Google Scholar]
  5. Brenner S., Stretton A. O., Kaplan S. Genetic code: the 'nonsense' triplets for chain termination and their suppression. Nature. 1965 Jun 5;206(988):994–998. doi: 10.1038/206994a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Burgess R. R., Travers A. A., Dunn J. J., Bautz E. K. Factor stimulating transcription by RNA polymerase. Nature. 1969 Jan 4;221(5175):43–46. doi: 10.1038/221043a0. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Fowler A. V., Zabin I. Co-linearity of beta-galactosidase with its gene by immunological detection of incomplete polypeptide chains. Science. 1966 Nov 25;154(3752):1027–1029. doi: 10.1126/science.154.3752.1027. [DOI] [PubMed] [Google Scholar]
  10. Jaskunas S. R., Burgess R. R., Nomura M. Identification of a gene for the alpha-subunit of RNA polymerase at the str-spc region of the Escherichia coli chromosome. Proc Natl Acad Sci U S A. 1975 Dec;72(12):5036–5040. doi: 10.1073/pnas.72.12.5036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Konrad B., Kirschbaum J., Austin S. Isolation and characterization of phi80 transducing bacteriophage for a ribonucleic acid polymerase gene. J Bacteriol. 1973 Nov;116(2):511–516. doi: 10.1128/jb.116.2.511-516.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  13. Newton A. Re-initiation of polypeptide synthesis and polarity in the lac operon of Escherichia coli. J Mol Biol. 1969 May 14;41(3):329–339. doi: 10.1016/0022-2836(69)90279-4. [DOI] [PubMed] [Google Scholar]
  14. STRETTON A. O., BRENNER S. MOLECULAR CONSEQUENCES OF THE AMBER MUTATION AND ITS SUPPRESSION. J Mol Biol. 1965 Jun;12:456–465. doi: 10.1016/s0022-2836(65)80268-6. [DOI] [PubMed] [Google Scholar]
  15. Wilcox G., Meuris P., Bass R., Englesberg E. Regulation of the L-arabinose operon BAD in vitro. J Biol Chem. 1974 May 10;249(9):2946–2952. [PubMed] [Google Scholar]
  16. Zubay G., Cheong L., Gefter M. DNA-directed cell-free synthesis of biologically active transfer RNA: su + 3 tyrosyl-tRNA. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2195–2197. doi: 10.1073/pnas.68.9.2195. [DOI] [PMC free article] [PubMed] [Google Scholar]

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