       Document 0639
 DOCN  M9640639
 TI    Construction of a type 1 human immunodeficiency virus that maintains a
       primer binding site complementary to tRNA(His).
 DT    9604
 AU    Wakefield JK; Kang SM; Morrow CD; Department of Microbiology, University
       of Alabama at Birmingham; 35294, USA.
 SO    J Virol. 1996 Feb;70(2):966-75. Unique Identifier : AIDSLINE
       MED/96135208
 AB    The initiation of human immunodeficiency virus type 1 reverse
       transcription occurs by extension of a tRNA(Lys3) primer bound near the
       5' end of the viral RNA genome which is designated the primer binding
       site (PBS). Sequences within the viral genome upstream of the PBS which
       are complementary to the anticodon loop (USUU) and the T psi C loop and
       arm (AGGGTm psi) of tRNA(Lys3) are postulated to play a role in
       maintaining the selective use of tRNA(Lys3) in reverse transcription. To
       investigate this possibility, proviral genomes which contain a PBS
       complementary to the 3'-terminal 18 nucleotides of tRNA(His)
       [pHXB2(His)] as well as sequences upstream of this PBS which are
       complementary to either the anticodon loop [CCACAA; pHXB2(His-AC)] or T
       psi C loop [GACCGAGG; pHXB2(His-T psi C)] of tRNA(His) were constructed.
       Infectious virus was recovered upon transfection into COS-1 cells of
       pHXB2(His), pHXB2(His-AC), or pHXB2(His-T psi C). The appearance of
       infectious virus after cocultivation with SupT1 cells was delayed for
       the proviruses containing a PBS complementary to tRNA(His) compared with
       that obtained by transfection of the wild-type provirus [pHXB2(WT)].
       However, by several passages in SupT1 cells, the mutant viruses
       demonstrated replication kinetics similar to those of the wild-type
       virus. A DNA sequence analysis of the PBS region from integrated
       proviruses revealed that by day 15 of culture, the PBS of viruses
       derived from pHXB2(His) and pHXB2(His-T psi C) reverted back to the
       wild-type PBS complementary to tRNA(Lys3). In contrast, viruses derived
       from pHXB2(His-AC) maintained a PBS complementary to tRNA(His) for over
       4 months in culture encompassing 12 serial passages. This study, then,
       is the first report of a stable human immunodeficiency virus type 1
       which utilizes an alternative tRNA primer and suggests that interactions
       between the primer tRNA anticodon loop and viral sequences upstream of
       the PBS contribute to the specificity of the tRNA primer used in reverse
       transcription.
 DE    Animal  Anticodon/GENETICS  Base Sequence  Binding Sites  Cell Line  DNA
       Primers  DNA, Viral/GENETICS/METABOLISM  Genome, Viral  Human
       HIV-1/*GENETICS  Molecular Sequence Data  Mutation  Nucleic Acid
       Conformation  Proviruses/GENETICS  RNA, Transfer, His/*METABOLISM  RNA,
       Transfer, Lys/GENETICS/METABOLISM  RNA, Viral/GENETICS/*METABOLISM
       Structure-Activity Relationship  Support, U.S. Gov't, P.H.S.
       Transcription, Genetic  Viral Proteins/METABOLISM  Virus Replication
       JOURNAL ARTICLE

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

