Abstract
Reverse transcription of the retroviral RNA genome begins with tRNA-primed synthesis of a minus-strand DNA, which subsequently acts as the template for the synthesis of plus-strand DNA. This plus-strand DNA is initiated at a unique location and makes use of a purine-rich RNA oligonucleotide derived by RNase H action on the viral RNA. To determine the variables that are relevant to successful specific initiation of plus-strand DNA synthesis, we have used nucleic acid sequences from the genome of Rous sarcoma virus along with three different sources of RNase H: avian myeloblastosis virus DNA polymerase, murine leukemia virus DNA polymerase, and the RNase H of Escherichia coli. Our findings include evidence that specificity is controlled not only by the nucleic acid sequences but also by the RNase H. For example, while the avian reverse transcriptase efficiently and specifically initiates on the sequences of the avian retrovirus, the murine reverse transcriptase initiates specifically but at a location 4 bases upstream of the correct site.
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.
- Champoux J. J., Gilboa E., Baltimore D. Mechanism of RNA primer removal by the RNase H activity of avian myeloblastosis virus reverse transcriptase. J Virol. 1984 Mar;49(3):686–691. doi: 10.1128/jvi.49.3.686-691.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen P. J., Cywinski A., Taylor J. M. Reverse transcription of 7S L RNA by an avian retrovirus. J Virol. 1985 May;54(2):278–284. doi: 10.1128/jvi.54.2.278-284.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gerard G. F., D'Alessio J. M., Kotewicz M. L., Noon M. C. Influence on stability in Escherichia coli of the carboxy-terminal structure of cloned Moloney murine leukemia virus reverse transcriptase. DNA. 1986 Aug;5(4):271–279. doi: 10.1089/dna.1986.5.271. [DOI] [PubMed] [Google Scholar]
- Kotewicz M. L., Sampson C. M., D'Alessio J. M., Gerard G. F. Isolation of cloned Moloney murine leukemia virus reverse transcriptase lacking ribonuclease H activity. Nucleic Acids Res. 1988 Jan 11;16(1):265–277. doi: 10.1093/nar/16.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makino S., Soe L. H., Shieh C. K., Lai M. M. Discontinuous transcription generates heterogeneity at the leader fusion sites of coronavirus mRNAs. J Virol. 1988 Oct;62(10):3870–3873. doi: 10.1128/jvi.62.10.3870-3873.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mason W. S., Taylor J. M., Hull R. Retroid virus genome replication. Adv Virus Res. 1987;32:35–96. doi: 10.1016/s0065-3527(08)60474-1. [DOI] [PubMed] [Google Scholar]
- Myers J. C., Dobkin C., Spiegelman S. RNA primer used in synthesis of anticomplementary DNA by reverse transcriptase of avian myeloblastosis virus. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1316–1320. doi: 10.1073/pnas.77.3.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olsen J. C., Watson K. F. Avian retrovirus RNA-directed DNA synthesis by purified reverse transcriptase. Covalent linkage of RNA to plus strand DNA. Biochem Biophys Res Commun. 1980 Dec 31;97(4):1376–1383. doi: 10.1016/s0006-291x(80)80019-2. [DOI] [PubMed] [Google Scholar]
- Omer C. A., Resnick R., Faras A. J. Evidence for involvement of an RNA primer in initiation of strong-stop plus DNA synthesis during reverse transcription in vitro. J Virol. 1984 May;50(2):465–470. doi: 10.1128/jvi.50.2.465-470.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Panganiban A. T., Temin H. M. The terminal nucleotides of retrovirus DNA are required for integration but not virus production. Nature. 1983 Nov 10;306(5939):155–160. doi: 10.1038/306155a0. [DOI] [PubMed] [Google Scholar]
- Rattray A. J., Champoux J. J. The role of Moloney murine leukemia virus RNase H activity in the formation of plus-strand primers. J Virol. 1987 Sep;61(9):2843–2851. doi: 10.1128/jvi.61.9.2843-2851.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Resnick R., Omer C. A., Faras A. J. Involvement of retrovirus reverse transcriptase-associated RNase H in the initiation of strong-stop (+) DNA synthesis and the generation of the long terminal repeat. J Virol. 1984 Sep;51(3):813–821. doi: 10.1128/jvi.51.3.813-821.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith J. K., Cywinski A., Taylor J. M. Initiation of plus-strand DNA synthesis during reverse transcription of an avian retrovirus genome. J Virol. 1984 Jan;49(1):200–204. doi: 10.1128/jvi.49.1.200-204.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith J. K., Cywinski A., Taylor J. M. Specificity of initiation of plus-strand DNA by Rous sarcoma virus. J Virol. 1984 Nov;52(2):314–319. doi: 10.1128/jvi.52.2.314-319.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sorge J., Hughes S. H. Polypurine tract adjacent to the U3 region of the Rous sarcoma virus genome provides a cis-acting function. J Virol. 1982 Aug;43(2):482–488. doi: 10.1128/jvi.43.2.482-488.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor J., Sharmeen L. Retrovirus genome replication: priming specificities of plus-strand DNA synthesis. J Cell Sci Suppl. 1987;7:189–195. doi: 10.1242/jcs.1987.supplement_7.14. [DOI] [PubMed] [Google Scholar]
- Verma I. M. Studies on reverse transcriptase of RNA tumor viruses III. Properties of purified Moloney murine leukemia virus DNA polymerase and associated RNase H. J Virol. 1975 Apr;15(4):843–854. doi: 10.1128/jvi.15.4.843-854.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]




