title: In virio SHAPE analysis of tRNA(Lys3) annealing to HIV-1 genomic RNA in wild type and protease-deficient virus.
abstract: BACKGROUND
  tRNA(Lys3) annealing to the viral RNA of human immunodeficiency virus type-1 (HIV-1) is an essential step in the virus life cycle, because this tRNA serves as the primer for initiating reverse transcription. tRNA(Lys3) annealing to viral RNA occurs in two steps. First, Gag promotes annealing of tRNA(Lys3) to the viral RNA during cytoplasmic HIV-1 assembly. Second, mature nucleocapsid (NCp7), produced from the processing of Gag by viral protease during viral budding from the cell, remodels the annealed complex to form a more stable interaction between the viral RNA and tRNA(Lys3), resulting in a more tightly bound and efficient primer for reverse transcription.
  
  
  RESULTS
  In this report, we have used in virio SHAPE analysis of both the 5´-untranslated region in HIV-1 RNA and the annealed tRNA(Lys3) to determine structural differences of the annealed complex that occur between protease-negative (Pr-) and wild type viruses. Our results indicate that the weaker binding of tRNA(Lys3) annealed by Gag in Pr- virions reflects both missing interactions of tRNA(Lys3) with viral RNA regions in the upper PBS stem, and a weaker interaction with the internal stem-loop found within the unannealed primer binding site in viral RNA.
  
  
  CONCLUSIONS
  We propose secondary structure models for the tRNA(Lys3)/viral RNA annealed complexes in PR- and wild type viruses that support the two-step annealing model by showing that Gag promotes a partial annealing of tRNA(Lys3) to HIV-1 viral RNA, followed by a more complete annealing by NCp7.
authors.affiliation: Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, Montreal, QC, H3T 1E2, Canada. seif.elias@gmail.com.
authors.forename: Elias
authors.initials: E
authors.lastname: Seif
authors.affiliation: Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, Montreal, QC, H3T 1E2, Canada. meijuann@yahoo.com.
authors.forename: Meijuan
authors.initials: M
authors.lastname: Niu
authors.affiliation: Lady Davis Institute for Medical Research and McGill AIDS Centre, Jewish General Hospital, Montreal, QC, H3T 1E2, Canada. lawrence.kleiman@mcgill.ca.|Department of Medicine, McGill University, Montreal, QC, H3A 1A1, Canada. lawrence.kleiman@mcgill.ca.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Kleiman
mesh_terms: D020121:5' Untranslated Regions; D016333:HIV Protease / Q000172:deficiency*; D015497:HIV-1 / Q000201:enzymology* / Q000502:physiology*; D006801:Humans; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012357:RNA, Transfer, Lys / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; C071884:NCP7 protein, Human immunodeficiency virus 1; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D054301:gag Gene Products, Human Immunodeficiency Virus; D016333:HIV Protease