       Document 0133
 DOCN  M9460133
 TI    Human immunodeficiency virus uses tRNA(Lys,3) as primer for reverse
       transcription in HeLa-CD4+ cells.
 DT    9404
 AU    Das AT; Koken SE; Essink BB; van Wamel JL; Berkhout B; Department of
       Virology, University of Amsterdam, The Netherlands.
 SO    FEBS Lett. 1994 Mar 14;341(1):49-53. Unique Identifier : AIDSLINE
       MED/94185771
 AB    Significant amounts of different tRNA molecules are present in
       retroviral particles, but one specific tRNA species functions as primer
       in reverse transcription. It is generally believed that the HIV-1 virus
       uses the tRNA(Lys,3) molecule as primer. This is based on sequence
       complementarity between the 3' end of tRNA(Lys,3) and the primer-binding
       site (PBS) on HIV-1 genomic RNA. Recent biochemical analyses indicated
       that tRNA(LYs,3) is indeed incorporated into viral particles.
       Interestingly, tRNA(Lys,3) could not be detected in virions produced by
       HeLa-CD4+ cells [(1992) Biochem. Biophys. Res. Commun. 185, 1105-1115].
       In order to test whether alternative tRNA molecules can function as
       primer in HIV replication, we performed a series of experiments based on
       the observation that tRNA primer sequences are inherited by the viral
       progeny. We cultured HIV-1 for prolonged periods of time in HeLa-CD4+
       cells, but did not detect sequence changes in the PBS region.
       Furthermore, we found PBS-mutants to be replication-incompetent, again
       suggesting that HIV-1 solely uses tRNA(Lys,3) as primer. Most
       importantly, we obtained revertants of one such PBS-mutant, which had
       restored a wild-type PBS sequence. This tRNA(Lys,3)-mediated repair
       demonstrates a general requirement for this primer in HIV-1 reverse
       transcription.
 DE    Antigens, CD4  Base Sequence  DNA, Viral  Hela Cells  Human
       HIV-1/ENZYMOLOGY/*GENETICS  Molecular Sequence Data  Reverse
       Transcriptase/*METABOLISM  RNA, Transfer, Lys/*METABOLISM  RNA,
       Viral/METABOLISM  Support, Non-U.S. Gov't  Transcription, Genetic
       Transfection  Virus Replication/PHYSIOLOGY  JOURNAL ARTICLE

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

