Document 0694
 DOCN  M9550694
 TI    Intracellular susceptibility to ribozymes in a tethered
       substrate-ribozyme provirus model is not predicted by secondary
       structures of human immunodeficiency virus type 1 RNAs in vitro.
 DT    9505
 AU    Dropulic B; Jeang KT; Molecular Virology Section, National Institute of
       Allergy and; Infectious Diseases, National Institutes of Health,
       Bethesda,; Maryland 20892.
 SO    Antisense Res Dev. 1994 Fall;4(3):217-21. Unique Identifier : AIDSLINE
       MED/95152268
 AB    We have assessed the sensitivity of different sites in HIV-1 genomic RNA
       to ribozymes. Ribozymes targeted to sequences in U5, Pol, Env, RRE, or R
       were positioned into nef of an infectious HIV-1 provirus. When these
       proviral DNAs were introduced into HeLa CD4+ cells, recombinant viruses
       that contain ribozymes tethered to genomic RNA or viral mRNAs were
       produced. The growth kinetics of ribozyme-containing viruses in CD4+
       lymphocytes (MT4 cells) were distinctly delayed when compared to control
       viruses. On the basis of the ability of a particular ribozyme to inhibit
       virus replication, we inferred intracellular ribozyme-sensitive sites.
       We found that although ribozyme sensitivity in vitro could be correlated
       with predicted secondary structures of target RNAs, such in vivo
       correlations could not be made when using the HIV provirus model. We
       conclude that both Zuker algorithm computer modeling of substrate RNA
       secondary structures and in vitro cleavage efficiencies cannot be
       reliably used to determine HIV-1 ribozyme sensitive sites in vivo.
 DE    Acquired Immunodeficiency Syndrome/ENZYMOLOGY  Chromosome Mapping
       Comparative Study  Hela Cells  Human  HIV-1/*GENETICS  Models,
       Biological  Nucleic Acid Conformation  *Proviruses  RNA,
       Catalytic/*METABOLISM  RNA, Viral/*GENETICS  Substrate Specificity
       Support, U.S. Gov't, P.H.S.  T-Lymphocytes/VIROLOGY  JOURNAL ARTICLE

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