Genetic assay for multimerization of retroviral gag polyproteins.

Abstract:

:We have established a genetic assay for the multimerization of retroviral gag polyproteins. This assay is based on the GAL4 two-hybrid system for studying protein-protein interactions (S. Fields and O. Song, Nature (London) 340:245-246, 1989). In our initial experiments, we generated Saccharomyces cerevisiae plasmids that separately express the GAL4 DNA-binding and GAL4 activation domains fused to the human immunodeficiency virus type 1 (HIV-1) gag polyprotein, Pr55gag. The coexpression of these two hybrid proteins in S. cerevisiae results in the association of the GAL4 domains and the potent activation of an integrated GAL4-responsive lacZ indicator gene. Similar results were obtained with plasmids encoding GAL4-Moloney murine leukemia virus (M-MuLV) gag polyprotein hybrid proteins. In contrast, the heterologous GAL4-HIV-1 gag and GAL4-M-MuLV gag fusion proteins were unable to interact with each other to induce lacZ expression. The results suggest that this yeast system provides a rapid and specific assay for the interactions of retroviral gag proteins that occur during virion assembly.

journal_name

J Virol

journal_title

Journal of virology

authors

Luban J,Alin KB,Bossolt KL,Humaran T,Goff SP

doi

10.1128/JVI.66.8.5157-5160.1992

subject

Has Abstract

pub_date

1992-08-01 00:00:00

pages

5157-60

issue

8

eissn

0022-538X

issn

1098-5514

journal_volume

66

pub_type

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