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M94A3295.TXT
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1994-10-25
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Document 3295
DOCN M94A3295
TI Internally initiated gag and gag-pol precursors of HIV-1.
DT 9412
AU Kojima A; Tachibana Y; Hoshikawa N; Kurata T; Department of Pathology,
National Institute of Health, Tokyo,; Japan.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):109 (abstract no. PA0053). Unique
Identifier : AIDSLINE ICA10/94369280
AB OBJECTIVE: Inspection of the gag ORF reveales a second methionin (M2)
codon which is 10 resides downstream of the p17-p24 junction. Our object
is to examine the de novo synthesis of internally initiated gag or
gag-pol precursors. METHODS: Mutant gag or gag-pol ORFs either lacking
the M2 codon, replaced by a leucine or isoleucine codon, or starting at
the M2 codon were constructed and expressed by vaccinia viruses. HIV-1
gag and pol proteins were analysed by western blotting, p24 ELISA and RT
assay. HIV-1-like particle formation was examined by sucrose density
centrifugation and electron microscopy. RESULTS AND DISCUSSION: (1)
Deletion or substitution of the M2 codon abolished the expression of a
40 kDa gag protein (Pr40gag) with no effects on the processing of
Pr55gag and Pr160gag-pol precursors. (2) M2-starting gag and gag-pol
ORFs expressed Pr40gag and Pr140gag-pol which did not generate mature
p24 nor RT activity. (3) The smaller precursors, Pr40gag and
Pr140gag-pol, lacked the ability to form HIV-like particles. These
results show the existence of internally initiated Pr40gag and
Pr140gag-pol polyprotein precursors. Involvement of the novel precursors
in the processing, assembling and packaging of the gag/pol proteins will
be discussed.
DE Codon/GENETICS Fusion Proteins, gag-pol/*BIOSYNTHESIS/GENETICS Gene
Products, gag/*BIOSYNTHESIS/GENETICS Genes, gag Genes, pol Human
HIV-1/GENETICS/*METABOLISM In Vitro Methionine/GENETICS Mutagenesis,
Site-Directed Open Reading Frames Protein
Precursors/*BIOSYNTHESIS/GENETICS Protein Processing,
Post-Translational Sequence Deletion Vaccinia Virus/GENETICS MEETING
ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).