       Document 0118
 DOCN  M9460118
 TI    Role of Pr160gag-pol in mediating the selective incorporation of
       tRNA(Lys) into human immunodeficiency virus type 1 particles.
 DT    9404
 AU    Mak J; Jiang M; Wainberg MA; Hammarskjold ML; Rekosh D; Kleiman L; Lady
       Davis Institute for Medical Research, Jewish General; Hospital,
       Montreal, Quebec, Canada.
 SO    J Virol. 1994 Apr;68(4):2065-72. Unique Identifier : AIDSLINE
       MED/94187044
 AB    COS-7 cells transfected with human immunodeficiency virus type 1 (HIV-1)
       proviral DNA produce virus in which three tRNA species are most abundant
       in the viral tRNA population. These tRNAs have been identified through
       RNA sequencing techniques as tRNA(3Lys) the primer tRNA in HIV-1, and
       members of the tRNA(1,2Lys) isoacceptor family. These RNAs represent 60%
       of the low-molecular-weight RNA isolated from virus particles, while
       they represent only 6% of the low-molecular-weight RNA isolated from the
       COS cell cytoplasm. Thus, tRNA(Lys) is selectively incorporated into
       HIV-1 particles. We have measured the ratio of tRNA(3Lys) molecules to
       copies of genomic RNA in viral RNA samples and have calculated that
       HIV-1 contains approximately eight molecules of tRNA(3Lys) per two
       copies of genomic RNA. We have also obtained evidence that the
       Pr160gag-pol precursor is involved in primer tRNA(3Lys) incorporation
       into virus. First, selective tRNA(Lys) incorporation and wild-type
       amounts of tRNA(3Lys) were maintained in a protease-negative virus
       unable to process Pr55gag and Pr160gag-pol precursors, indicating that
       precursor processing was not required for primer tRNA incorporation.
       Second, viral particles containing only unprocessed Pr55gag protein did
       not selectively incorporate tRNA(Lys), while virions containing both
       unprocessed Pr55gag and Pr160gag-pol proteins demonstrated select
       tRNA(3Lys) packaging. Third, studies with a proviral mutant containing a
       deletion of most of the reverse transcriptase sequences and
       approximately one-third of the integrase sequence in the Pr160gag-pol
       precursor resulted in the loss of selective tRNA incorporation and an
       eightfold decrease in the amount of tRNA(3Lys) per two copies of genomic
       RNA. We have also confirmed herein finding of a previous study which
       indicated that the primer binding site is not required for the selective
       incorporation of tRNA(Lys).
 DE    Base Sequence  Fusion Proteins, gag-pol/GENETICS/*METABOLISM  Gene
       Products, gag/METABOLISM  HIV Protease/GENETICS/METABOLISM
       HIV-1/*GROWTH & DEVELOPMENT/GENETICS  Molecular Sequence Data
       Polymerase Chain Reaction  Protein Precursors/METABOLISM  Protein
       Processing, Post-Translational  Proviruses/*GROWTH &
       DEVELOPMENT/GENETICS  Reverse Transcriptase/METABOLISM  RNA  RNA,
       Transfer, Lys/*METABOLISM  RNA, Viral/ANALYSIS  Support, Non-U.S. Gov't
       Support, U.S. Gov't, P.H.S.  Transcription, Genetic  Virion/*GROWTH &
       DEVELOPMENT/GENETICS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

