Champoux, James J. and Gilboa, Eli and Baltimore, David (1984) Mechanism of RNA primer removal by the RNase H activity of avian myeloblastosis virus reverse transcriptase. Journal of Virology, 49 (3). pp. 686-691. ISSN 0022-538X. http://resolver.caltech.edu/CaltechAUTHORS:CHAjvir84
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The single-stranded DNA containing the Moloney murine leukemia virus origin for plus-strand synthesis was cloned in M13mp2 and used as a template for avian myeloblastosis virus reverse transcriptase in the presence of Moloney RNA which had been treated with pancreatic RNase A. The RNA pieces containing the polypurine stretch near the plus-strand origin were processed, presumably by RNase H, to generate primers for DNA synthesis which initiated both at the correct origin site and at one nucleotide downstream from the correct site. Approximately 50% of the labeled DNA fragments synthesized under these conditions retained the priming RNA on their 5' ends. When the isolated fragments were hybridized back to the template DNA and again treated with the reverse transcriptase, all of the RNA was removed from the labeled DNA. By using 5'-end-labeled pancreatic RNase A-resistant fragments, it was possible to show that the RNA primers were removed intact. It appears from these results that the RNase H activity associated with the enzyme shows a preference for cutting at the junction between the RNA and DNA moieties of such complexes and therefore is ideally suited for removing RNA primers.
|Additional Information:||Copyright © 1984 by the American Society for Microbiology. Received 16 September 1983/Accepted 6 December 1983 We thank Michael Been for his critical reading of the manuscript. This work was supported by Public Health Service grant CA 26717 from the National Cancer Institute and in part by grant no. NP-400 from the American Cancer Society. D.B. is an American Cancer Society Professor of Microbiology. J.J.C. was supported by a John Simon Guggenheim Memorial Foundation Fellowship.|
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|Deposited On:||26 Apr 2006|
|Last Modified:||26 Dec 2012 08:50|
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