Footprint Analysis of Replicating Murine Leukemia Virus Reverse Transcriptase
- 6 January 1995
- journal article
- other
- Published by American Association for the Advancement of Science (AAAS) in Science
- Vol. 267 (5194) , 96-99
- https://doi.org/10.1126/science.7528942
Abstract
Replication complexes that contained either murine leukemia virus reverse transcriptase (MLV RT) or a variant reverse transcriptase without a ribonuclease (RNase) H domain (delta RH MLV RT) were visualized by enzymatic footprinting. Wild-type MLV RT protected template nucleotides +6 to -27, and primer nucleotides -1 to -26 of primers that had first been extended by one or four nucleotides. Although it catalyzed DNA synthesis, delta RH MLV RT stably bound template-primer only under conditions of reduced ionic strength and protected the duplex portion only as far as position -15. Despite altered hydrolysis profiles, both enzymes covered primarily the template-primer duplex, contradicting recent predictions based on the structure of rat DNA polymerase beta.Keywords
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