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. 1981 Apr;78(4):2057–2061. doi: 10.1073/pnas.78.4.2057

Analysis of avian myeloblastosis viral RNA and in vitro synthesis of proviral DNA.

R A Schulz, J G Chirikjian, T S Papas
PMCID: PMC319282  PMID: 6264460

Abstract

Two virus-specific RNA species of 7.5 and 7.0 kilobases have been identified in avian myeloblastosis virus (AMV) by denaturing gel electrophoresis and blot hybridization analysis, and they were found to be in a 10:1 ratio. The individual RNAs direccted the cell-free synthesis of the 76,000-dalton "gag" protein and the 180,000-dalton "gal-pol" protein, thereby demonstrating 5' sequence homology of approximately 4.9 kilobases between the two species. Synthesis of these two precursor proteins by the AMV genome indicates structural differences between AMV and other avian acute leukemia viruses. The two viral RNAs were transcribed into complete cDNA copies with AMV DNA polymerase. Linear proviruses were found to be 90--100% resistant to S1 nuclease. Analysis of single-stranded transcripts demonstrated two distinct species of 2.6 and 2.3 x 10(6) daltons, and analysis of duplexes formed from the single-stranded transcripts demonstrated species of 5.2 and 4.0 x 10(6) daltons.

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Selected References

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

  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bister K., Duesberg P. H. Genetic structure of avian acute leukemia viruses. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):801–822. doi: 10.1101/sqb.1980.044.01.086. [DOI] [PubMed] [Google Scholar]
  3. Bister K., Duesberg P. H. Genetic structure of avian acute leukemia viruses. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):801–822. doi: 10.1101/sqb.1980.044.01.086. [DOI] [PubMed] [Google Scholar]
  4. Clayman C. H., Mosharrafa E., Faras A. J. In vitro synthesis of infectious transforming DNA by the avian sarcoma virus reverse transcriptase. J Virol. 1979 Jan;29(1):242–249. doi: 10.1128/jvi.29.1.242-249.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fritsch E., Temin H. M. Formation and structure of infectious DNA of spleen necrosis virus. J Virol. 1977 Jan;21(1):119–130. doi: 10.1128/jvi.21.1.119-130.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gilboa E., Mitra S. W., Goff S., Baltimore D. A detailed model of reverse transcription and tests of crucial aspects. Cell. 1979 Sep;18(1):93–100. doi: 10.1016/0092-8674(79)90357-x. [DOI] [PubMed] [Google Scholar]
  7. Hughes S. H., Shank P. R., Spector D. H., Kung H. J., Bishop J. M., Varmus H. E., Vogt P. K., Breitman M. L. Proviruses of avian sarcoma virus are terminally redundant, co-extensive with unintegrated linear DNA and integrated at many sites. Cell. 1978 Dec;15(4):1397–1410. doi: 10.1016/0092-8674(78)90064-8. [DOI] [PubMed] [Google Scholar]
  8. Kacian D. L., Myers J. C. Synthesis of extensive, possibly complete, DNA copies of poliovirus RNA in high yields and at high specific activities. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2191–2195. doi: 10.1073/pnas.73.7.2191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Lai M. M., Hu S. S. In vitro synthesis and characterisation of full- and half-genome length complementary DNA from avian oncoviruses. Nature. 1978 Feb 2;271(5644):481–483. doi: 10.1038/271481a0. [DOI] [PubMed] [Google Scholar]
  11. Langridge J., Langridge P., Bergquist P. L. Extraction of nucleic acids from agarose gels. Anal Biochem. 1980 Apr;103(2):264–271. doi: 10.1016/0003-2697(80)90266-3. [DOI] [PubMed] [Google Scholar]
  12. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  13. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  14. Moscovici C. Leukemic transformation with avian myeloblastosis virus: present status. Curr Top Microbiol Immunol. 1975;71:79–101. doi: 10.1007/978-3-642-66193-8_2. [DOI] [PubMed] [Google Scholar]
  15. Moscovici C., Vogt P. K. Effects of genetic cellular resistance on cell transformation and virus replication in chicken hematopoietic cell cultures infected with avian myeloblastosis virus (BAI-A). Virology. 1968 Aug;35(4):487–497. doi: 10.1016/0042-6822(68)90278-x. [DOI] [PubMed] [Google Scholar]
  16. Myers J. C., Spiegelman S., Kacian D. L. Synthesis of full-length DNA copies of avian myeloblastosis virus RNA in high yields. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2840–2843. doi: 10.1073/pnas.74.7.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Paterson B. M., Marciani D. J., Papas T. S. Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4951–4954. doi: 10.1073/pnas.74.11.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Stephenson M. L., Wirthlin L. S., Scott J. F., Zamecnik P. C. The 3'-terminal nucleosides of the high molecular weight RNA of avian myeloblastosis virus. Proc Natl Acad Sci U S A. 1972 May;69(5):1176–1180. doi: 10.1073/pnas.69.5.1176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Westphal H., Lai S. P. Quantitative electron microscopy of early adenovirus RNA. J Mol Biol. 1977 Nov 5;116(3):525–548. doi: 10.1016/0022-2836(77)90082-1. [DOI] [PubMed] [Google Scholar]

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