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M94A3331.TXT
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Document 3331
DOCN M94A3331
TI Functional analysis of HIV-1 glycoproteins with deletions in gp120.
DT 9412
AU Adams O; Schaal H; Scheid A; Universitat Dusseldorf, Germany.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):100 (abstract no. PA0020). Unique
Identifier : AIDSLINE ICA10/94369244
AB OBJECTIVES: (1) To determine the contribution of gp120 to the membrane
fusion activity of the HIV-1 glycoprotein and (2) to determine the
structured requirements for HIV-1 glycoprotein oligomerization. METHODS:
A vector was constructed which expresses the HIV-1 env, tat, and rev
genes under the control of the SV40 early promotor. From this clone two
vectors were derived expressing a glycoprotein lacking all but the
C-terminal 10 and 50 amino acid residues of gp120 and two further
derivatives expressing N-terminal sequences of the rat vasopressin
precursor protein. Processing of mutant Env-proteins was monitored in
HeLa T4+ and in COS cells. Fusion activity was monitored after
transfection in HeLa T4+ cells and transdominant interference was
measured after co-transfection of a wild-type gp120 vector with the
deletion- and substitution mutants. RESULTS AND CONCLUSIONS: Mutant
proteins with gp120 truncations and substitutions were processed in COS
cells resulting in the formation of gp41. In HeLa T4+ cells, cleavage
products of truncated env-genes were smaller than gp41 and could not be
not safely identified with vasopressin hybrid proteins. While
transfection of HeLa T4+ cells with wild type env vectors results in
extensive cell-cell fusion, no giant cell formation could be detected
after transfection with truncated or substituted gp120 mutants. Thus, we
could not find syncytium formation with gp41 alone. This is in contrast
to findings with similiar constructs (J. Virol. 66, 4134, 1992).
Co-transfection of cells with both wt-env vectors and gp120 mutants
resulted in inhibition of cell-cell fusion indicating that glycoprotein
oligomerization may occur with mutant glycoproteins lacking most of
gp120. This effect was more pronounced with vasopressin mutants.
DE Animal Cell Line Cloning, Molecular Genetic Vectors Hela Cells
Human HIV Envelope Protein gp120/*GENETICS/*PHYSIOLOGY HIV Envelope
Protein gp41/PHYSIOLOGY HIV-1/*GENETICS/*PHYSIOLOGY Membrane
Fusion/GENETICS/PHYSIOLOGY Protein Processing, Post-Translational Rats
Recombinant Fusion Proteins/GENETICS *Sequence Deletion Transfection
Vasopressins/GENETICS MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).