Document 0317
 DOCN  M9590317
 TI    Stepwise analysis of reverse transcription in a cell-to-cell HIV
       infection model: kinetics and implications.
 DT    9509
 AU    Karageorgos L; Li P; Burrell CJ; NCHVR, Division of Medical Virology,
       IMVS, Adelaide SA.
 SO    Annu Conf Australas Soc HIV Med. 1994 Nov 3-6;6:186 (unnumbered
       abstract). Unique Identifier : AIDSLINE ASHM6/95291782
 AB    We have investigated the kinetics of human immunodeficiency virus (HIV)
       reverse transcription in infected T-cells, using a synchronized,
       one-step, cell-to-cell infection model and quantitative PCR assays for
       the different DNA intermediate structures that are found sequentially
       during reverse transcription. After an initial lag period, the minus
       strand strong-stop viral DNA was first detected 1.5 hr after the
       initiation of cell-to-cell HIV infection. The post-transfer, newly
       extended minus strand viral DNA was first detected 2 hr post infection,
       whereas both plus strand strong-stop DNA and fully extended minus strand
       DNA were first detected at 2.5 hr post infection. Kinetic data suggested
       that once the reverse transcription was initiated, the HIV reverse
       transcriptase synthesized minus strand DNA at a rate of 150-180 bases
       per minute, and that the first template transfer and the initiation of
       the plus strand DNA synthesis imposed specific time delays of 15-30
       minutes and 40-50 minutes respectively. In contrast, minus strand viral
       DNA synthesized after the second template transfer appeared at a time
       point very close to the time of the appearance of the last piece of DNA
       synthesized just before the second template switch suggesting that the
       second switch occurred very rapidly. Taken together, our results define
       more accurately than previously possible the rates of several of the
       sequential steps in HIV reverse transcription in infected T-cell lines.
       Our findings also implicate different mechanisms for the two distinct
       template switches during retrovirus reverse transcription. Finally, this
       study demonstrates the use of PCR to quantitate DNA intermediates that
       may be heterogeneous in length but define a population of molecules that
       have completed a discrete step during a one-step growth cycle.
 DE    Cell Transformation, Viral/GENETICS  Cells, Cultured  DNA,
       Viral/GENETICS  Human  HIV Infections/*TRANSMISSION/VIROLOGY
       HIV-1/*GENETICS  Reverse Transcriptase/*GENETICS
       T-Lymphocytes/*VIROLOGY  Templates  Virus Integration/GENETICS  Virus
       Replication/*GENETICS  MEETING ABSTRACT

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