Abstract
RNA polymerase was precipitated from extracts of radioactively labeled vegetative and sporulating Bacillus subtilis with antiserum prepared against vegetative core polymerase. The precipitates were solubilized and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antiserum added to an extract of vegetative B. subtilis precipitated only the known subunits of core RNA polymerase, but antiserum added to an extract of sporulating cells precipitated a new polypeptide of 70,000 daltons in addition to the subunits of core enzyme. The 70,000-dalton polypeptide precipitated from an extract of a mixture of vegetative and sporulating B. subtilis, separately labeled with two different radioisotopes, contained only the radioisotope characteristic of the sporulating cells.
The 70,000-dalton protein has been freed of core RNA polymerase and extensively purified by chromatography on phosphocellulose. Precipitation of the purified 70,000-dalton protein by the anti-polymerase serum requires the prior addition of vegetative or sporulation core RNA polymerase. The reaction is specific since the purified protein is not precipitated during antibody precipitation of either phage λ repressor or bovine serum albumin.
The RNA polymerase-binding protein appears during the third hour of sporulation and is apparently not synthesized by the sporulation-defective mutant rfr 10.
Keywords: antibody precipitation
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Hussey C., Losick R., Sonenshein A. L. Ribosomal RNA synthesis is turned off during sporulation of Bacillus subtilis. J Mol Biol. 1971 Apr 14;57(1):59–70. doi: 10.1016/0022-2836(71)90119-7. [DOI] [PubMed] [Google Scholar]
- Hussey C., Pero J., Shorenstein R. G., Losick R. In vitro synthesis of ribosomal RNA by Bacillus subtilis RNA polymerase. Proc Natl Acad Sci U S A. 1972 Feb;69(2):407–411. doi: 10.1073/pnas.69.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kornberg A., Spudich J. A., Nelson D. L., Deutscher M. P. Origin of proteins in sporulation. Annu Rev Biochem. 1968;37:51–78. doi: 10.1146/annurev.bi.37.070168.000411. [DOI] [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]
- Losick R., Shorenstein R. G., Sonenshein A. L. Structural alteration of RNA polymerase during sporulation. Nature. 1970 Aug 29;227(5261):910–913. doi: 10.1038/227910a0. [DOI] [PubMed] [Google Scholar]
- Losick R., Sonenshein A. L. Change in the template specificity of RNA polymerase during sporulation of Bacillus subtilis. Nature. 1969 Oct 4;224(5214):35–37. doi: 10.1038/224035a0. [DOI] [PubMed] [Google Scholar]
- Richardson J. P. RNA polymerase and the control of RNA synthesis. Prog Nucleic Acid Res Mol Biol. 1969;9:75–116. doi: 10.1016/s0079-6603(08)60768-0. [DOI] [PubMed] [Google Scholar]
- 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]
- Sonenshein A. L., Roscoe D. H. The course of phage phi-e infection in sporulating cells of Bacillus subtilis strain 3610. Virology. 1969 Oct;39(2):265–275. doi: 10.1016/0042-6822(69)90047-6. [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]