title: Preferences for the selection of unique tRNA primers revealed from analysis of HIV-1 replication in peripheral blood mononuclear cells.
abstract: BACKGROUND
  All human immunodeficiency virus (HIV-1) uses a host tRNALys,3 as the primer for reverse transcription. The tRNALys,3 is bound to a region on the HIV-1 genome, the primer-binding site (PBS), that is complementary to the 18 terminal nucleotides of tRNALys,3. How HIV-1 selects the tRNA from the intracellular milieu is unresolved.
  
  
  RESULTS
  HIV-1 tRNA primer selection has been investigated using viruses in which the primer-binding site (PBS) and a sequence within U5 were altered so as to be complementary to tRNAMet, tRNAPro or tRNAIle. Analysis of the replication of these viruses in human peripheral blood mononuclear cells (PBMC) revealed preferences for the selection of certain tRNAs. HIV-1 with the PBS altered to be complementary to tRNAMet, with and without the additional mutation in U5 to be complementary to the anticodon of tRNAMet, stably maintains the PBS complementary to tRNAMet following extended in vitro culture in PBMC. In contrast, viruses with either the PBS or PBS and U5 mutated to be complementary to tRNAIle were unstable during in vitro replication in PBMC and reverted to utilize tRNALys,3. Viruses with the PBS altered to be complementary to tRNAPro replicated in PBMC but reverted to use tRNALys,3; viruses with mutations in both the U5 and PBS complementary to tRNAPro maintained this PBS, yet replicated poorly in PBMC.
  
  
  CONCLUSIONS
  The results of these studies demonstrate that HIV-1 has preferences for selection of certain tRNAs for high-level replication in PBMC.
authors.affiliation: Department of Cell Biology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA. kendalee@uab.edu
authors.forename: Kenda L
authors.initials: KL
authors.lastname: Moore-Rigdon
authors.forename: Barry R
authors.initials: BR
authors.lastname: Kosloff
authors.forename: Richard L
authors.initials: RL
authors.lastname: Kirkman
authors.forename: Casey D
authors.initials: CD
authors.lastname: Morrow
mesh_terms: D001665:Binding Sites; D017931:DNA Primers; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology*; D009154:Mutation; D012345:RNA, Transfer, Amino Acid-Specific / Q000235:genetics / Q000378:metabolism*; D012357:RNA, Transfer, Lys / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D017415:Ribonucleoprotein, U5 Small Nuclear / Q000235:genetics; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D012345:RNA, Transfer, Amino Acid-Specific; D012357:RNA, Transfer, Lys; D012367:RNA, Viral; D017415:Ribonucleoprotein, U5 Small Nuclear; D054303:HIV Reverse Transcriptase