title: Nuclease footprinting of human immunodeficiency virus reverse transcriptase/tRNA(Lys-3) complexes.
abstract: Nuclease footprinting has been used to probe features of binary complexes of type 1 human immunodeficiency virus reverse transcriptase (HIV-1 RT) with both natural and synthetic preparations of its cognate replication primer, tRNA(Lys-3). In addition to heterodimeric RT (p66/p51), ribonucleoprotein complexes containing either the p66 or p51 subunit were analyzed. Footprinting experiments employed both structure- and sequence-specific nucleases. Our results indicate a similar mode of interaction for the three RT preparations tested, suggesting contact with each loop of the tRNA primer (D, anticodon, and T psi C), as well as minor perturbation of the anticodon stem. Although there is little evidence for extensive disruption of the 3'-acceptor stem. RNase A footprinting data with natural and synthetic tRNA suggests that potential base pairing between the T psi C and D loops is disrupted in the presence of RT.
authors.affiliation: Division of Infectious Diseases, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106-4984.
authors.forename: B M
authors.initials: BM
authors.lastname: Wöhrl
authors.forename: B
authors.initials: B
authors.lastname: Ehresmann
authors.forename: G
authors.initials: G
authors.lastname: Keith
authors.forename: S F
authors.initials: SF
authors.lastname: Le Grice
mesh_terms: D000926:Anticodon; D001483:Base Sequence; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012330:RNA, Double-Stranded / Q000378:metabolism; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012259:Ribonuclease, Pancreatic / Q000378:metabolism
chemical_list: D000926:Anticodon; D012330:RNA, Double-Stranded; D012357:RNA, Transfer, Lys; D011994:Recombinant Proteins; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase; D012259:Ribonuclease, Pancreatic