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M9480625.TXT
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1994-08-20
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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).