Document 0505
 DOCN  M9490505
 TI    Preferential incorporation of nucleoside analogs after template
       switching during human immunodeficiency virus reverse transcription.
 DT    9411
 AU    Arts EJ; Wainberg MA; McGill AIDS Centre, Lady Davis Institute-Jewish
       General Hospital,; Montreal, Quebec, Canada.
 SO    Antimicrob Agents Chemother. 1994 May;38(5):1008-16. Unique Identifier :
       AIDSLINE MED/94346798
 AB    We assessed the effects of 3'-azido-3'-deoxythymidine (AZT),
       2',3'-dideoxyinosine (ddI), and the (-) enantiomer of
       2',3'-dideoxy-3'-thiacytidine (3TC) on reverse transcription in
       CD4-positive cells by isolating truncated human immunodeficiency virus
       (HIV) DNA fragments. Jurkat cells were treated with AZT (2 microM), ddI
       (200 microM), or 3TC (50 microM) prior to infection with HIV.
       Low-molecular-weight DNA was isolated and amplified by PCR with primer
       pairs which identify different segments of HIV proviral DNA. We found
       that the HIV DNA fragments generated from drug-treated, HIV-exposed
       Jurkat cells were truncated at a ratio of 15:1 [i.e., (-) strong-stop
       DNA to HIV DNA generated after the first template switch]. Full-length
       DNA was observed in the case of untreated, HIV-infected cultures.
       Following nucleoside analog treatment of HIV-exposed Jurkat cells,
       reverse transcription was terminated only after the synthesis of (-)
       strong-stop DNA. The nucleoside analogs tested, i.e., AZT, ddI, and 3TC,
       preferentially chain terminated viral DNA synthesis immediately
       following the first template switch. The (-) strong-stop HIV DNA was
       present in AZT-treated and untreated cultures for at least 6 days. We
       also carried out cell-free reverse transcription/template-switching
       reactions involving tRNA(Lys3) or a deoxyoligonucleotide as a primer, as
       a means of studying the selective incorporation of AZT triphosphate into
       proviral DNA. When reactions were primed with tRNA(Lys3), we found that
       AZT triphosphate was preferentially incorporated after template
       switching.
 DE    Antiviral Agents/METABOLISM  Base Sequence  Cell Line
       Didanosine/METABOLISM  DNA, Viral/ANALYSIS/ISOLATION & PURIF  Human
       HIV-1/GENETICS/*METABOLISM  Molecular Sequence Data
       Nucleosides/*METABOLISM  Polymerase Chain Reaction  RNA, Transfer,
       Lys/METABOLISM  Support, Non-U.S. Gov't  Templates  Thymine
       Nucleotides/METABOLISM  *Transcription, Genetic  T4 Lymphocytes/DRUG
       EFFECTS/METABOLISM/MICROBIOLOGY  Zidovudine/ANALOGS &
       DERIVATIVES/METABOLISM  JOURNAL ARTICLE

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