PMID: 6162482Jan 20, 1981Paper

Mechanism of action of Moloney murine leukemia virus RNA-directed DNA polymerase associated RNase H (RNase H I)

Biochemistry
G F Gerard

Abstract

The mechanism of action of the ribonuclease H (RNase H) activity associated with Moloney murine leukemia virus RNA-directed DNA polymerase (RNase H I) and the two-subunit (alpha beta) form of avian myeloblastosis virus DNA polymerase were compared by utilizing the model substrate (A)n.(dT)n and polyacrylamide gel electrophoresis in 7 M urea to analyze digestion products. Examination on 25% polyacrylamide gels revealed that a larger proportion of the RNase H I oligonucleotide products generated by limited digestion of [3H](A)(1100).(dT)n were acid insoluble (15-26 nucleotides long) than acid soluble (less than 15 nucleotides long), while the opposite was true for products generated by alpha beta RNase H. RNase H I was capable of attacking RNA in RNA.DNA in the 5' to 3' and 3' to 5' directions, as demonstrated by the use of [3H,3'- or 5'-32P](A)(380).(dT)n and cellulose--[3H](A)n.(dT)n. Both RNase H I and alpha beta RNase H degraded [3H]-(A)n.(dT)n with a partially processive mechanism, based upon classical substrate competition experiments and analyses of the kinetics of degradation of [3H,3'- or 5'-32P](A)(380).(dT)n. That is, both enzymes remain bound to a RNA.DNA substrate through a finite number of hydrolytic events but diss...Continue Reading

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Citations

Jan 1, 1990·Pharmacology & Therapeutics·U Wintersberger
Jun 1, 1997·Bioorganic & Medicinal Chemistry·F TakusagawaR F Weaver
May 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·M S Krug, S L Berger
Oct 1, 1981·European Journal of Biochemistry·C FrelinM Lazdunski
Jan 1, 1992·Faraday Discussions·H JungblutH J Lewerenz
Jun 27, 1997·Journal of Molecular Biology·G W HanR E Dickerson
Jul 1, 1989·Applied and Environmental Microbiology·R Weller, D M Ward

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