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. 1974 Aug;14(2):249–260. doi: 10.1128/jvi.14.2.249-260.1974

Characterization of Late Polyoma mRNA

Elena Buetti a,1
PMCID: PMC355510  PMID: 4367901

Abstract

Polyoma-infected mouse kidney cell cultures were labeled with [3H]uridine for 3 h late in the lytic cycle (26 to 29 h after infection) and RNA was extracted from cytoplasm and nuclei and from isolated polyribosomes. Sedimentation velocity analysis in sucrose gradients showed that polyoma-specific “giant” and 26S RNAs are present exclusively in the nucleus. RNA associated with cytoplasmic polyribosomes was analyzed by sedimentation in aqueous sucrose density gradients and dimethylsulfoxide sucrose gradients, as well as by polyacrylamide gel electrophoresis. Polyoma-specific RNA in polyribosomes consists of at least two classes, with sedimentation coefficients of 16 (major fraction) and 19S (minor fraction) in aqueous sucrose gradients and 15 and 17S, respectively, in dimethylsulfoxide gradients. Estimates based on dimethylsulfoxide gradient and analysis suggest a molecular weight of approximately 500,000 for 16S RNA and 700,000 for 19S RNA. These polyoma RNAs seem to undergo reversible conformational changes under the different conditions of analysis. We cannot exclude the possibility that they contain more than one molecular species.

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Selected References

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

  1. Acheson N. H., Buetti E., Scherrer K., Weil R. Transcription of the polyoma virus genome: synthesis and cleavage of giant late polyoma-specific RNA. Proc Natl Acad Sci U S A. 1971 Sep;68(9):2231–2235. doi: 10.1073/pnas.68.9.2231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borun T. W., Scharff M. D., Robbins E. Preparation of mammalian polyribosomes with the detergent Nonidet P-40. Biochim Biophys Acta. 1967 Nov 21;149(1):302–304. doi: 10.1016/0005-2787(67)90715-0. [DOI] [PubMed] [Google Scholar]
  3. Friedmann T. Genetic economy of polyoma virus: capsid proteins are cleavage products of same viral gene. Proc Natl Acad Sci U S A. 1974 Feb;71(2):257–259. doi: 10.1073/pnas.71.2.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grollman A. P. Structural basis for inhibition of protein synthesis by emetine and cycloheximide based on an analogy between ipecac alkaloids and glutarimide antibiotics. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1867–1874. doi: 10.1073/pnas.56.6.1867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kajioka R. Some studies on the status of polyoma-specified RNA after transcription. Virology. 1972 Apr;48(1):284–287. doi: 10.1016/0042-6822(72)90140-7. [DOI] [PubMed] [Google Scholar]
  6. Kolakofsky D., Boy de la Tour E., Delius H. Molecular weight determination of Sendai and Newcastle disease virus RNA. J Virol. 1974 Feb;13(2):261–268. doi: 10.1128/jvi.13.2.261-268.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Martin M. A., Axelrod D. Polyoma virus gene activity during lytic infection and in transformed animal cells. Science. 1969 Apr 4;164(3875):68–70. doi: 10.1126/science.164.3875.68. [DOI] [PubMed] [Google Scholar]
  8. May E., May P., Weil R. "Early" virus-specific RNA may contain information necessary for chromosome replication and mitosis induced by Simian Virus 40. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1654–1658. doi: 10.1073/pnas.70.6.1654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. McConkey E. H., Hopkins J. W. Molecular weights of some HeLa ribosomal RNA's. J Mol Biol. 1969 Feb 14;39(3):545–550. doi: 10.1016/0022-2836(69)90144-2. [DOI] [PubMed] [Google Scholar]
  10. Mirault M. E., Scherrer K. Isolation of preribosomes from HeLa cells and their characterization by electrophoresis on uniform and exponential-gradient-polyacrylamide gels. Eur J Biochem. 1971 Nov 11;23(2):372–386. doi: 10.1111/j.1432-1033.1971.tb01631.x. [DOI] [PubMed] [Google Scholar]
  11. Pene J. J., Knight E., Jr, Darnell J. E., Jr Characterization of a new low molecular weight RNA in HeLa cell ribosomes. J Mol Biol. 1968 May 14;33(3):609–623. doi: 10.1016/0022-2836(68)90309-4. [DOI] [PubMed] [Google Scholar]
  12. Penman S., Vesco C., Penman M. Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J Mol Biol. 1968 May 28;34(1):49–60. doi: 10.1016/0022-2836(68)90234-9. [DOI] [PubMed] [Google Scholar]
  13. Perry R. P., Kelley D. E. Messenger RNA-protein complexes and newly synthesized ribosomal subunits: analysis of free particles and components of polyribosomes. J Mol Biol. 1968 Jul 14;35(1):37–59. doi: 10.1016/s0022-2836(68)80035-x. [DOI] [PubMed] [Google Scholar]
  14. Perry R. P., La Torre J., Kelley D. E., Greenberg J. R. On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim Biophys Acta. 1972 Mar 14;262(2):220–226. doi: 10.1016/0005-2787(72)90236-5. [DOI] [PubMed] [Google Scholar]
  15. Petric M., Hudson J. B. The processing of polyoma virus RNA synthesized during productive infection in mouse cells. Can J Biochem. 1972 Aug;50(8):927–935. doi: 10.1139/o72-129. [DOI] [PubMed] [Google Scholar]
  16. Pétursson G., Weil R. A study on the mechanism of polyoma-induced activation of the cellular DNA-synthesizing apparatus. Synchronization by FUdR of virus-induced DNA synthesis. Arch Gesamte Virusforsch. 1968;24(1):1–29. doi: 10.1007/BF01242898. [DOI] [PubMed] [Google Scholar]
  17. Roblin R., Härle E., Dulbecco R. Polyoma virus proteins. 1. Multiple virion components. Virology. 1971 Sep;45(3):555–566. doi: 10.1016/0042-6822(71)90171-1. [DOI] [PubMed] [Google Scholar]
  18. Seehafer J. G., Weil R. Synthesis of polyoma virus structural polypeptides in mouse kidney cells. Virology. 1974 Mar;58(1):75–85. doi: 10.1016/0042-6822(74)90142-1. [DOI] [PubMed] [Google Scholar]
  19. Strauss J. H., Jr, Kelly R. B., Sinsheimer R. L. Denaturation of RNA with dimethyl sulfoxide. Biopolymers. 1968 Jun;6(6):793–807. doi: 10.1002/bip.1968.360060604. [DOI] [PubMed] [Google Scholar]
  20. Vaughan M. H., Warner J. R., Darnell J. E. Ribosomal precursor particles in the HeLa cell nucleus. J Mol Biol. 1967 Apr 28;25(2):235–251. doi: 10.1016/0022-2836(67)90140-4. [DOI] [PubMed] [Google Scholar]
  21. WINOCOUR E. Purification of polyoma virus. Virology. 1963 Feb;19:158–168. doi: 10.1016/0042-6822(63)90005-9. [DOI] [PubMed] [Google Scholar]
  22. Weil R., Kára J. Polyoma "tumor antigen": an activator of chromosome replication? Proc Natl Acad Sci U S A. 1970 Oct;67(2):1011–1017. doi: 10.1073/pnas.67.2.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weinberg R. A., Warnaar S. O., Winocour E. Isolation and characterization of simian virus 40 ribonucleic acid. J Virol. 1972 Aug;10(2):193–201. doi: 10.1128/jvi.10.2.193-201.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]

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