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Nucleic Acids Research, 1995, Vol. 23, No. 15 2811-2814
© 1995

RNA

Reverse transcriptase reads through a 2'–5' linkage and a 2'-thiphosphate in a template

John R. Lorsch, David P. Bartel+ and Jack W. Szostak*

Department of Molecular Biology, Massachusetts General Hospital Boston, MA 02114, USA

*To whom correspondence should be addressed

Received June 16, 1995. Accepted June 27, 1995.

Avian myeloblastosis virus and Maloney murlne leukemia virus RNase H– reverse transcriptases pause when they encounter a 2'–5' linkage or a 2'-thiophosphate In their template RNAs, but eventually read through these backbone modifications. Both reverse transcriptases pause after the 2'–5' linkage but before the 2'-thlophosphate. These results suggest that in the absence of precise information concerning the behavior of a given reverse transcriptase with respect to a particular lesion or modification, caution should be exercised in the interpretation of primer extension data that is being used to determine the existence of, or map the position of, a crosslink, site of chemical modification or non-standard linkage in an RNA template.

+Present address: Whitehead Institute for Biomedical Research, Cambridge, MA 04, USA


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