Abstract
The enzyme, Qβ replicase, responsible for the replication of the RNA of Escherichia coli pahge Qβ, is composed of four nonidentical subunits, three of which, I, III, and IV, are coded for by the bacterial genome, while subunit II is phage-specific.
Subunit IV is shown to be identical to the protein synthesis elongation factor EF Ts by the following criteria: coelectrophoresis on polyacrylamide gels in sodium dodecyl sulfate and in urea buffers, identity of the first seven amino acids at the amino-terminus, precipitation of sub-unit IV by anti-EF T-factor serum, and stimulation of EF Tu-GDP exchange by subunit IV. Subunit III is shown to be identical to the protein synthesis elongation factor EF Tu by the following criteria: coelectrophoresis on sodium dodecyl sulfate gels, precipitation of EF Tu by anti-Qβ replicase serum, binding of guanine nucleotides, and binding of phenylalanyl-tRNA. In addition, Qβ replicase activity can be reconstituted from subunits I and II with EF Tu and EF Ts.
Keywords: RNA phage, transfer factors, ppGpp, renaturation
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beaud G., Lengyel P. Peptide chain elongation. Role of the S 1 factor in the pathway from S 3 -guanosine diphosphate complex to aminoacyl transfer ribonucleic acid-S 3 -guanosine triphosphate complex. Biochemistry. 1971 Dec 21;10(26):4899–4906. doi: 10.1021/bi00802a011. [DOI] [PubMed] [Google Scholar]
- Cashel M., Gallant J. Two compounds implicated in the function of the RC gene of Escherichia coli. Nature. 1969 Mar 1;221(5183):838–841. doi: 10.1038/221838a0. [DOI] [PubMed] [Google Scholar]
- Edlin G., Broda P. Physiology and genetics of the "ribonucleic acid control" locus in escherichia coli. Bacteriol Rev. 1968 Sep;32(3):206–226. doi: 10.1128/br.32.3.206-226.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fedoroff N. V., Zinder N. D. Structure of the poly(G) polymerase component of the bacteriophage f2 replicase. Proc Natl Acad Sci U S A. 1971 Aug;68(8):1838–1843. doi: 10.1073/pnas.68.8.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goodman H. M., Billeter M. A., Hindley J., Weissmann C. The nucleotide sequence at the 5'-terminus of the Q RNA minus trand. Proc Natl Acad Sci U S A. 1970 Oct;67(2):921–928. doi: 10.1073/pnas.67.2.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gordon J., Weissbach H. Immunochemical distinction between the Escherichia coli polypeptide chain elongation factors Tu and Ts. Biochemistry. 1970 Oct 13;9(21):4233–4236. doi: 10.1021/bi00823a028. [DOI] [PubMed] [Google Scholar]
- Haseltine W. A. In vitro transcription of Escherichia coli ribosomal RNA genes. Nature. 1972 Feb 11;235(5337):329–333. doi: 10.1038/235329a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Kamen R. Characterization of the subunits of Q-beta replicase. Nature. 1970 Nov 7;228(5271):527–533. doi: 10.1038/228527a0. [DOI] [PubMed] [Google Scholar]
- Kondo M., Gallerani R., Weissmann C. Subunit structure of Q-beta replicase. Nature. 1970 Nov 7;228(5271):525–527. doi: 10.1038/228525a0. [DOI] [PubMed] [Google Scholar]
- Lockwood A. H., Hattman S., Dubnoff J. S., Maitra U. Formation and properties of T factor complexes. J Biol Chem. 1971 May 10;246(9):2936–2947. [PubMed] [Google Scholar]
- Lucas-Lenard J., Lipmann F. Separation of three microbial amino acid polymerization factors. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1562–1566. doi: 10.1073/pnas.55.6.1562. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lucas-Lenard J. Protein biosynthesis. Annu Rev Biochem. 1971;40:409–448. doi: 10.1146/annurev.bi.40.070171.002205. [DOI] [PubMed] [Google Scholar]
- Pettijohn D. E. Ordered and preferential initiation of ribosomal RNA synthesis in vitro. Nat New Biol. 1972 Feb 16;235(59):204–206. doi: 10.1038/newbio235204a0. [DOI] [PubMed] [Google Scholar]
- Stavis R. L., August J. T. The biochemistry of RNA bacteriophage replication. Annu Rev Biochem. 1970;39:527–560. doi: 10.1146/annurev.bi.39.070170.002523. [DOI] [PubMed] [Google Scholar]
- Travers A. A., Kamen R. I., Schleif R. F. Factor necessary for ribosomal RNA synthesis. Nature. 1970 Nov 21;228(5273):748–751. doi: 10.1038/228748a0. [DOI] [PubMed] [Google Scholar]
- Weber K., Kuter D. J. Reversible denaturation of enzymes by sodium dodecyl sulfate. J Biol Chem. 1971 Jul 25;246(14):4504–4509. [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]
- Weissbach H., Miller D. L., Hachmann J. Studies on the role of factor Ts in polypeptide synthesis. Arch Biochem Biophys. 1970 Mar;137(1):262–269. doi: 10.1016/0003-9861(70)90433-9. [DOI] [PubMed] [Google Scholar]
- Weissbach H., Redfield B., Brot N. Aminoacyl-tRNA-Tu-GTP interaction with ribosomes. Arch Biochem Biophys. 1971 Aug;145(2):676–684. doi: 10.1016/s0003-9861(71)80028-0. [DOI] [PubMed] [Google Scholar]
- de Wachter R., Merregaert J., Vandenberghe A., Contreras R., Fiers W. Studies on the bacteriophage MS2. The untranslated 5'-terminal nucleotide sequence preceding the first cistron. Eur J Biochem. 1971 Oct 14;22(3):400–414. doi: 10.1111/j.1432-1033.1971.tb01558.x. [DOI] [PubMed] [Google Scholar]