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M9490075.TXT
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Document 0075
DOCN M9490075
TI Initiation of reverse transcription during cell-to-cell transmission of
human immunodeficiency virus infection uses pre-existing reverse
transcriptase.
DT 9411
AU Li P; Stephenson AJ; Brennan PA; Karageorgos L; Kok T; Kuiper LJ; Swift
J; Burrell CJ; National Centre for HIV Virology Research, Institute of
Medical; and Veterinary Science, Adelaide, Australia.
SO J Gen Virol. 1994 Aug;75 ( Pt 8):1917-26. Unique Identifier : AIDSLINE
MED/94321981
AB H3B cells, a laboratory clone of H9 cells persistently infected with the
HTLV-IIIB strain of human immunodeficiency virus (HIV), contained
significant levels of cell-associated reverse transcriptase (RT)
activity measured by in vitro assays using either exogenous or
endogenous templates. The cell-associated RT activity detected using
exogenous template was almost wholly in a soluble (non-sedimentable)
form whereas endogenous activity sedimented as a particulate structure
associated with viral RNA. Despite this, H3B cells did not contain
episomal HIV DNA detectable by Southern blot, indicating that in vivo
reverse transcription was not occurring to any significant extent in
these cells. However, when susceptible HUT 78 cells were infected by
co-cultivation with H3B cells, dramatic synthesis of episomal HIV DNA
occurred. Concurrently with this de novo initiation of reverse
transcription, however, we found no detectable change in intracellular
levels or cleavage profiles of immunoprecipitable RT polypeptides.
Finally, actinomycin D pre-treatment of H3B cells to prevent de novo
transcription from donor cell proviral DNA after co-cultivation did not
affect the initiation of in vivo reverse transcription following
cell-to-cell HIV infection. These results demonstrated that cells
persistently infected with HIV contained significant fully cleaved
cell-associated RT in a form that was active in vitro but not in vivo
and that following cell-to-cell transmission of HIV infection to
susceptible cells, de novo reverse transcription was initiated without
detectable evidence of further synthesis or proteolytic processing of
HIV RT. The nature of this initiation process requires further study.
DE Biological Transport Blotting, Western Cells, Cultured Human
HIV/ENZYMOLOGY/*GROWTH & DEVELOPMENT/GENETICS/ULTRASTRUCTURE
Microscopy, Electron Precipitin Tests Reverse
Transcriptase/*METABOLISM RNA, Viral/ISOLATION & PURIF Support,
Non-U.S. Gov't *Transcription, Genetic Virus Integration JOURNAL
ARTICLE
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).