Document 0625
 DOCN  M9480625
 TI    Inhibition of human immunodeficiency virus type-1 env expression by C-5
       propyne oligonucleotides specific for Rev-response element stem-loop V.
 DT    9410
 AU    Fenster SD; Wagner RW; Froehler BC; Chin DJ; Agouron Institute, La
       Jolla, California 92037.
 SO    Biochemistry. 1994 Jul 19;33(28):8391-8. Unique Identifier : AIDSLINE
       MED/94304852
 AB    The binding of Rev to the Rev-response element (RRE) of the human
       immunodeficiency virus (HIV) is essential for RNA transport and
       expression of structural proteins such as gp160 encoded by env. To
       determine if env expression could be disrupted by complementary
       oligodeoxynucleotides (ODNs), band-shift studies were used to identify
       RRE sites that are essential for the formation of Rev-RRE complexes
       [Chin, D. J. (1992) J. Virol. 66, 600-607] or the stability of preformed
       complexes. In this report, we describe complete disruption of preformed
       Rev-RRE complexes by a subset of 15 ODNs complementary to stem-loop V.
       The most potent ODN complementary to bases CUGGGGCAUCAAGC disrupted 50%
       of preformed complexes at 1.2 microM, a 400-fold molar excess over the
       RNA. Expression of env in COS7 cells was blocked by nuclear
       microinjection of ODNs with C-5 propyne-modified pyrimidines and
       phosphorothioate linkages. Inhibition was highly dependent upon RNA
       target position, internucleotide chemistry, ODN sequence, and
       concentration. Unmodified phosphodiester or phosphorothioate ODNs were
       inactive. For the most potent ODN, 50% of the injected cells' env
       expression (I50) was blocked with 0.1 microM. A translational block is
       unlikely since these ODNs blocked expression of a luciferase vector in
       which the RRE was placed downstream of the termination codon. Consistent
       with their in vitro effects upon Rev-RRE complexes, stem-loop V ODNs
       were 9-fold more active than stem-loop II ODNs in blocking env
       expression while having a reduced (I50 = 0.27 microM) but equivalent
       potency against luciferase-RRE. These results suggest that disruption of
       Rev-RRE complexes may assist in blocking env expression.
 DE    beta-Galactosidase/GENETICS  Alkynes/*PHARMACOLOGY  Base Sequence
       Binding Sites  Cell Line  DNA, Complementary/PHARMACOLOGY  DNA,
       Viral/CHEMISTRY/METABOLISM  Gene Expression/*DRUG EFFECTS  Gene
       Products, env/GENETICS/METABOLISM  Gene Transfer  *Genes, env  *Genes,
       rev  HIV-1/*GENETICS  Microinjections  Molecular Sequence Data  Nucleic
       Acid Conformation  Oligodeoxyribonucleotides/*PHARMACOLOGY  Protein
       Precursors/GENETICS/METABOLISM  Ribonuclease H, Calf Thymus/METABOLISM
       Support, U.S. Gov't, P.H.S.  JOURNAL ARTICLE

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