Abstract
E. coli B cells infected with T3 phage contain a modified host RNA polymerase in addition to the normal RNA polymerase found in uninfected cells. The modified RNA polymerase behaves differently in its elution properties from the normal enzyme on DEAE-cellulose, phosphocellulose, and DNA-cellulose column chromatography. The modified enzyme also differs from the normal polymerase in some of its enzymatic parameters. The specific activity of the modified RNA polymerase is markedly lower (i.e., ¼) than that of the normal enzyme. The decrease in activity is probably due to an alteration in the β′ subunit of the polymerase. A similar modification is also observed in nonpermissive cells infected with a gene-l amber mutant incapable of producing active T3 specific polymerase. An analogous modification in host RNA polymerase does not seem to occur in E. coli cells infected with T7 phage.
Keywords: DEAE-cellulose, phosphocellulose, and DNA-cellulose chromatography; β′ subunit
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- Burgess R. R. A new method for the large scale purification of Escherichia coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Biol Chem. 1969 Nov 25;244(22):6160–6167. [PubMed] [Google Scholar]
- Burgess R. R. RNA polymerase. Annu Rev Biochem. 1971;40:711–740. doi: 10.1146/annurev.bi.40.070171.003431. [DOI] [PubMed] [Google Scholar]
- Dunn J. J., McAllister W. T., Bautz E. K. Transcription in vitro of T3 DNA by Escherichia coli and T3 RNA polymerases. Analysis of the products in cell-free protein-synthesizing system. Eur J Biochem. 1972 Sep 25;29(3):500–508. doi: 10.1111/j.1432-1033.1972.tb02014.x. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Losick R. In vitro transcription. Annu Rev Biochem. 1972;41:409–446. doi: 10.1146/annurev.bi.41.070172.002205. [DOI] [PubMed] [Google Scholar]
- Mahadik S. P., Dharmgrongartama B., Srinivasan P. R. An inhibitory protein of Escherichia coli RNA polymerase in bacteriophage T3-infected cells (core polymerase-sigma factor-host polymerase-phage polymerase-initiation). Proc Natl Acad Sci U S A. 1972 Jan;69(1):162–166. doi: 10.1073/pnas.69.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maitra U., Huang H. H. Initiation, release, and reinitiation of RNA chains by bacteriophage-T3-induced polymerase from T3 DNA templates (E. coli-guanosine triphosphate terminus-purified polymerase). Proc Natl Acad Sci U S A. 1972 Jan;69(1):55–59. doi: 10.1073/pnas.69.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schweiger M., Herrlich P., Scherzinger E., Rahmsdorf H. J. Negative control of protein synthesis after infection with bacteriophage T7. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2203–2207. doi: 10.1073/pnas.69.8.2203. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sethi V. S. Structure and function of DNA-dependent RNA-polymerase. Prog Biophys Mol Biol. 1971;23:67–101. doi: 10.1016/0079-6107(71)90017-4. [DOI] [PubMed] [Google Scholar]
- Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
- Studier F. W. Bacteriophage T7. Science. 1972 Apr 28;176(4033):367–376. doi: 10.1126/science.176.4033.367. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]