Enzymatic activities associated with avian and murine retroviral DNA polymerases. Catalysis of and active site involvement in pyrophosphate exchange and pyrophosphorolysis reactions
- PMID: 6153389
Enzymatic activities associated with avian and murine retroviral DNA polymerases. Catalysis of and active site involvement in pyrophosphate exchange and pyrophosphorolysis reactions
Abstract
Reverse transcriptase isolated from avian myeloblastosis virus (AMV) and Rauscher murine leukemia virus (RLV) were examined for their ability to catalyze polymerization, ribonuclease H, pyrophosphate exchange, and pyrophosphorolysis reactions. A detailed characterization and a study of requirements for the expression of pyrophosphate exchange and pyrophosphorolysis reactions indicated that a variety of RNA and DNA template-primers supported these catalytic reactions. Furthermore, hydrogen bonding of template to primer was essential, although RNA:RNA template-primers, e.g. poly(rA) . (rU)9 or 70 S RNA . tRNA complex, were not utilized for these reactions. AMV enzyme required Mg2+, and RLV enzyme Mn2+, as the preferred divalent metal ion for the expression of these activities. Response of various catalytic reactions to site-specific inhibitors revealed that polymerization and pyrophosphate exchange reactions were susceptible to reagents that affected either the substrate or the template binding site, intrinsic zinc, or sulfhydryl groups. RNase H and pyrophosphorolysis activities, on the other hand, exhibited susceptibility only to the template site-specific reagent. We, therefore, conclude that RNase H and pyrophosphorolysis reactions are catalyzed through the template binding site while polymerization and pyrophosphate exchange reactions require additional participation of the substrate binding site, as well as that of intrinsic zinc and the presence of reactive sulfhydryl groups.
Similar articles
-
On the fidelity of DNA replication. Enzyme activities associated with DNA polymerases from RNA tumor viruses.J Biol Chem. 1976 Feb 25;251(4):975-81. J Biol Chem. 1976. PMID: 55414
-
Comparative biochemical properties of RNA-directed DNA polymerases from Rauscher murine leukemia virus and avian myeloblastosis virus.Cancer Res. 1974 Oct;34(10):2585-93. Cancer Res. 1974. PMID: 4137585 No abstract available.
-
Reverse transcriptase-associated RNase H activity. II. Inhibition by natural and synthetic RNA.J Virol. 1978 Sep;27(3):576-81. doi: 10.1128/JVI.27.3.576-581.1978. J Virol. 1978. PMID: 81313 Free PMC article.
-
Observations on template-specific conditions for DNA synthesis by avian myeloblastosis virus DNA polymerase.Nucleic Acids Res. 1976 Jun;3(6):1473-86. doi: 10.1093/nar/3.6.1473. Nucleic Acids Res. 1976. PMID: 60740 Free PMC article.
-
Model RNA-directed DNA synthesis by avian myeloblastosis virus DNA polymerase and its associated RNase H.Biochemistry. 1979 Jul 24;18(15):3210-9. doi: 10.1021/bi00582a004. Biochemistry. 1979. PMID: 88956
Cited by
-
Amino acid sequence analysis of reverse transcriptase subunits from avian myeloblastosis virus.J Virol. 1980 Oct;36(1):115-9. doi: 10.1128/JVI.36.1.115-119.1980. J Virol. 1980. PMID: 6160262 Free PMC article.
-
Efficient pyrophosphorolysis by a hepatitis B virus polymerase may be a primer-unblocking mechanism.Proc Natl Acad Sci U S A. 2001 Apr 24;98(9):4984-9. doi: 10.1073/pnas.091324398. Proc Natl Acad Sci U S A. 2001. PMID: 11320247 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
