title: Footprint analysis of replicating murine leukemia virus reverse transcriptase.
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.
authors.affiliation: Division of Infectious Diseases, Case Western Reserve University School of Medicine, Cleveland, OH 44106.
authors.forename: B M
authors.initials: BM
authors.lastname: Wöhrl
authors.forename: M M
authors.initials: MM
authors.lastname: Georgiadis
authors.forename: A
authors.initials: A
authors.lastname: Telesnitsky
authors.forename: W A
authors.initials: WA
authors.lastname: Hendrickson
authors.forename: S F
authors.initials: SF
authors.lastname: Le Grice
mesh_terms: D004247:DNA / Q000096:biosynthesis; D017931:DNA Primers / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D054303:HIV Reverse Transcriptase; D006868:Hydrolysis; D009052:Leukemia Virus, Murine / Q000201:enzymology*; D012194:RNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D016914:Ribonuclease H / Q000378:metabolism; D013698:Templates, Genetic
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D004247:DNA; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D003850:Deoxyribonuclease I; D016914:Ribonuclease H