title: Structural constraints in the HIV-1 reverse transcriptase-primer/template complex for the initiation of DNA synthesis from primer tRNALys3.
abstract: The topography and functional implications of the complex formed in vitro between human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) and its primer tRNALys3 were studied in this work. On the basis of previous results showing the high affinity both of the native primer, tRNALys3, as well as that of mismatched short oligonucleotide primers for HIV-1 RT, we synthesized chimeric primers containing tRNALys3 linked to U and T residues of different lengths. We found that the affinity of the oligonucleotide primers for HIV-1 RT is dramatically increased when linked to primer tRNA. Our results also show that in the tRNA.RT complex, before annealing tRNALys3 to the retroviral RNA genome, the 3'-terminal nucleotide of tRNALys3 is positioned at a distance of one nucleotide unit away from the template in the active polymerization site of the enzyme.
authors.affiliation: Novosibirsk Institute of Bioorganic Chemistry, Russian Academy of Sciences, Novosibirsk.
authors.forename: O D
authors.initials: OD
authors.lastname: Zakharova
authors.forename: I V
authors.initials: IV
authors.lastname: Martiyanov
authors.forename: O A
authors.initials: OA
authors.lastname: Timofeeva
authors.forename: T I
authors.initials: TI
authors.lastname: Kolocheva
authors.forename: M
authors.initials: M
authors.lastname: Fournier
authors.forename: M L
authors.initials: ML
authors.lastname: Andreola
authors.forename: G A
authors.initials: GA
authors.lastname: Maksakova
authors.forename: V I
authors.initials: VI
authors.lastname: Yamkovoy
authors.forename: S
authors.initials: S
authors.lastname: Litvak
authors.forename: L
authors.initials: L
authors.lastname: Tarrago-Litvak
authors.forename: G A
authors.initials: GA
authors.lastname: Nevinsky
mesh_terms: D001665:Binding Sites; D003596:Cytosine / Q000737:chemistry; D004279:DNA, Viral / Q000138:chemical synthesis* / Q000737:chemistry; D054303:HIV Reverse Transcriptase / Q000737:chemistry*; D007700:Kinetics; D009841:Oligonucleotides / Q000138:chemical synthesis; D011061:Poly A / Q000737:chemistry; D012313:RNA / Q000737:chemistry*; D012357:RNA, Transfer, Lys / Q000138:chemical synthesis / Q000737:chemistry*; D013698:Templates, Genetic; D013941:Thymine / Q000737:chemistry
chemical_list: D004279:DNA, Viral; D009841:Oligonucleotides; C034477:RNA primers; D012357:RNA, Transfer, Lys; D011061:Poly A; D012313:RNA; D003596:Cytosine; D054303:HIV Reverse Transcriptase; D013941:Thymine