title: Structures of complexes formed by HIV-1 reverse transcriptase at a termination site of DNA synthesis.
abstract: This study presents structural parameters associated with termination of human immunodeficiency virus, type 1 (HIV-1) reverse transcriptase (RT) at Ter2, the major termination site located in the center of the HIV-1 genome. DNA footprinting studies of various elongation complexes formed by RT around wild type and mutant Ter2 sites have revealed two major structural transformations of these complexes when the enzyme gets closer to Ter2. First, the interactions between RT and the DNA duplex are less extended, although the global affinity of the enzyme for this duplex is only decreased by 2-fold. Second, there is an atypical positioning of the RT RNase H domain on the DNA duplex. We interpret our data as indicating that the A(n)T(m) motif located upstream of Ter2 prevents a classical positioning of the enzyme on the double-stranded part of the DNA duplex at some precise positions of elongation downstream of this motif. Instead, novel species of binary and/or ternary complexes, characterized by atypical footprints, are formed. The new rate-limiting step of the reaction, characterized in the preceding paper (Lavigne, M., Polomack, L., and Buc, H. (2001) J. Biol. Chem. 276, 31429-31438), would be a transition leading from these new species to a catalytically competent ternary complex.
authors.affiliation: Unité de Physicochimie des Macromolécules Biologiques, Institut Pasteur, CNRS URA 1773, 75724 Paris Cedex 15, France. lavigne@molbio.mgh.harvard.edu
authors.forename: M
authors.initials: M
authors.lastname: Lavigne
authors.forename: L
authors.initials: L
authors.lastname: Polomack
authors.forename: H
authors.initials: H
authors.lastname: Buc
mesh_terms: D001483:Base Sequence; D004279:DNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D003845:Deoxycytosine Nucleotides / Q000494:pharmacology; D054303:HIV Reverse Transcriptase / Q000737:chemistry*; D007700:Kinetics; D008969:Molecular Sequence Data; D016914:Ribonuclease H / Q000494:pharmacology
chemical_list: D004279:DNA, Viral; D003845:Deoxycytosine Nucleotides; C024107:2'-deoxycytidine 5'-triphosphate; D054303:HIV Reverse Transcriptase; D016914:Ribonuclease H