Abstract
Much work has been done on the isolation, purification, and characterization of the RNA-directed RNA polymerase (EC 2.7.7.48) of cucumber mosaic virus (CMV)-infected cucumbers. Uninfected plants were reported to have no such enzyme, but we recently detected low levels of the activity in cucumber. Since tobacco and cowpea contain such an enzyme that is variably increased in amount by various virus (as well as viroid) infections, we assumed that this would also be the case upon CMV infection of cucumber. However, further purification and characterization of the RNA-directed RNA polymerases from healthy and from infected cucumber suggests that these are different enzymes. The presumed CMV replicase was obtained pure and consists of a major polypeptide of Mr 100,000 and minor components of Mr 110,000 and about 10,000. The Km is 5 microM ([3H]GTP) when tobacco mosaic virus RNA is used as template.
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- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Clerx C. M., Bol J. F. Properties of solubilized RNA-dependent RNA polymerase from alfalfa mosaic virus-infected and healthy tobacco plants. Virology. 1978 Dec;91(2):453–463. doi: 10.1016/0042-6822(78)90391-4. [DOI] [PubMed] [Google Scholar]
- Fraenkel-Conrat H. RNA polymerase from tobacco necrosis virus infected and uninfected tobacco. Purification of the membrane-associated enzyme. Virology. 1976 Jul 1;72(1):23–32. doi: 10.1016/0042-6822(76)90308-1. [DOI] [PubMed] [Google Scholar]
- Fraenkel-Conrat H. RNA-dependent RNA polymerases of plants. Proc Natl Acad Sci U S A. 1983 Jan;80(2):422–424. doi: 10.1073/pnas.80.2.422. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franssen H., Leunissen J., Goldbach R., Lomonossoff G., Zimmern D. Homologous sequences in non-structural proteins from cowpea mosaic virus and picornaviruses. EMBO J. 1984 Apr;3(4):855–861. doi: 10.1002/j.1460-2075.1984.tb01896.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hadidi A., Fraenkel-Conrat H. Characterization and specificity of soluble RNA polymerase of brome mosaic virus. Virology. 1973 Apr;52(2):363–372. doi: 10.1016/0042-6822(73)90331-0. [DOI] [PubMed] [Google Scholar]
- Hardy S. F., German T. L., Loesch-Fries L. S., Hall T. C. Highly active template-specific RNA-dependent RNA polymerase from barley leaves infected with brome mosaic virus. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4956–4960. doi: 10.1073/pnas.76.10.4956. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ikegami M., Fraenkel-Conrat H. Characterization of the RNA-dependent RNA polymerase of tobacco leaves. J Biol Chem. 1979 Jan 10;254(1):149–154. [PubMed] [Google Scholar]
- Khan Z. A., Fraenkel-Conrat H. Purification and characterization of polynucleotide phosphorylase from cucumber. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1311–1315. doi: 10.1073/pnas.82.5.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kumarasamy R., Symons R. H. Extensive purification of the cucumber mosaic virus-induced RNA replicase. Virology. 1979 Jul 30;96(2):622–632. doi: 10.1016/0042-6822(79)90118-1. [DOI] [PubMed] [Google Scholar]
- 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]
- Le Roy C., Stussi-Garaud C., Hirth L. RNA-dependent RNA polymerases in uninfected and in alfalfa mosaic virus-infected tobacco plants. Virology. 1977 Oct 1;82(1):48–62. doi: 10.1016/0042-6822(77)90031-9. [DOI] [PubMed] [Google Scholar]
- Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
- Romaine C. P., Zaitlin M. RNA-dependent RNA polymerases in uninfected and tobacco mosaic virus-infected tabacco leaves: viral induced stimulation of a host polymerase activity. Virology. 1978 May 1;86(1):241–253. doi: 10.1016/0042-6822(78)90024-7. [DOI] [PubMed] [Google Scholar]
- Stussi-Garaud C., Lemius J., Fraenkel-Conrat H. RNA polymerase from tobacco necrosis virus-infected and uninfected tobacco. II. Properties of the bound and soluble polymerases and the nature of their products. Virology. 1977 Sep;81(2):224–236. doi: 10.1016/0042-6822(77)90139-8. [DOI] [PubMed] [Google Scholar]
- Takanami Y., Fraenkel-Conrat H. Comparative studies on ribonucleic acid dependent RNA polymerases in cucumber mosaic virus infected cucumber and tobacco and uninfected tobacco plants. Biochemistry. 1982 Jun 22;21(13):3161–3167. doi: 10.1021/bi00256a020. [DOI] [PubMed] [Google Scholar]
- White J. L., Dawson W. O. Characterization of RNA-dependent RNA polymerases in uninfected and cowpea chlorotic mottle virus-infected cowpea leaves: selective removal of host RNA polymerase from membranes containing CCMV RNA replicase. Virology. 1978 Jul 1;88(1):33–43. doi: 10.1016/0042-6822(78)90107-1. [DOI] [PubMed] [Google Scholar]