Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1976 May;3(5):1167–1183. doi: 10.1093/nar/3.5.1167

Isolation and characterization of poly(A)-containing intranuclear polyoma-specific "giant" RNA'S.

L J Rosenthal, C Salomon, R Weil
PMCID: PMC342978  PMID: 181731

Abstract

Heterogeneous polyoma giant RNA molecules have been isolated by oligo(dT)- cellulose chromatography during the late phase of a lytic cycle of infection of mouse kidney cell cultures. These RNAs have sedimentation coefficients in denaturing Me2SO gradients that are greater than 26S and thus apparently correspond to RNA molecules larger than one strand of polyoma DNA. Approximately 15% of total nuclear polyoma late giant RNAs contained tracts of poly(A) and were retained by oligo(dT)-cellulose. The polyoma late giant RNAs as well as heterogeneous nuclear RNAs (HnRNAs) were found to have a slightly lower sedimentation rate in Me2SO-chloral hydrate density gradients than sedimentation values in sucrose gradients indicated. Even when synthesis of viral DNA and the production of capsid protein are blocked by 5-fluorodeoxyuridine (FdU), 10% of polyoma-specific RNA (as determined by sedimentation analyses under aqueous conditions) was shown to contain tracts of poly(A). In contrast to our findings on polyoma late giant RNA, nuclear polyoma RNA synthesized in the presence of FdU sedimented in denaturing Me2SO-chloral hydrate gradients considerably slower (from 15 to 30S) in relation to HnRNA and ribosomal precursor RNA. The sedimentation pattern in denaturing Me2SO gradients suggest that Py RNA synthesized late in lytic infection in the presence of FdU may be no longer than one transcript of Py DNA.

Full text

PDF
1167

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. Buetti E. Characterization of late polyoma mRNA. J Virol. 1974 Aug;14(2):249–260. doi: 10.1128/jvi.14.2.249-260.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Darnell J. E., Jr Ribonucleic acids from animal cells. Bacteriol Rev. 1968 Sep;32(3):262–290. doi: 10.1128/br.32.3.262-290.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Edmonds M., Vaughan M. H., Jr, Nakazato H. Polyadenylic acid sequences in the heterogeneous nuclear RNA and rapidly-labeled polyribosomal RNA of HeLa cells: possible evidence for a precursor relationship. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1336–1340. doi: 10.1073/pnas.68.6.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Greenberg J. R., Perry R. P. Relative occurrence of polyadenylic acid sequences in messenger and heterogeneous nuclear RNA of L cells as determined by poly (U)-hydroxylapatite chromatography. J Mol Biol. 1972 Dec 14;72(1):91–98. doi: 10.1016/0022-2836(72)90070-8. [DOI] [PubMed] [Google Scholar]
  6. Hancock R., Weil R. Biochemical evidence for induction by polyoma virus of replication of the chromosomes of mouse kidney cells. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1144–1150. doi: 10.1073/pnas.63.4.1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kirby K. S., Cook E. A. Isolation of deoxyribonucleic acid from mammalian tissues. Biochem J. 1967 Jul;104(1):254–257. doi: 10.1042/bj1040254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kára J., Weil R. Specific activation of the DNA-synthesizing apparatus in contact-inhibited mouse kidney cells by polyoma virus. Proc Natl Acad Sci U S A. 1967 Jan;57(1):63–70. doi: 10.1073/pnas.57.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Parish J. H., Kirby K. S. Reagents which reduce interactions between ribosomal RNA and rapidly labelled RNA from rat liver. Biochim Biophys Acta. 1966 Dec 21;129(3):554–562. doi: 10.1016/0005-2787(66)90070-0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. WINOCOUR E. Purification of polyoma virus. Virology. 1963 Feb;19:158–168. doi: 10.1016/0042-6822(63)90005-9. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Weil R., Michel M. R., Ruschmann G. K. Induction of cellular DNA synthesis by polyoma virus. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1468–1475. doi: 10.1073/pnas.53.6.1468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Weil R., Salomon E., May E., May P. A simplifying concept in tumor virology: virus-specific "pleiotropic effectors". Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):381–395. doi: 10.1101/sqb.1974.039.01.050. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES