Liu LX, Benichou S, Schwartz O, Benarous R; Conference on Retroviruses and Opportunistic Infections.
Program Abstr 4th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 4th 1997 Wash DC. 1997 Jan 22-26; 4th: 202 (abstract no. 743).
INSERM, Paris, France.
We used the yeast two-hybrid system to identify cellular proteins that interact with HIV-1 Nef and could act as mediators of Nef function. In addition to beta-COP which was previously described, we have isolated cDNA encoding a new type of thioesterase displaying 42% identity with thioesterase II from E. coli, an enzyme involved in cleavage of thioester bonds. Indeed we found that the recombinant protein has thioesterase enzymatic activity. The interaction between Nef and thioesterase was confirmed both in vitro using recombinant proteins and in cells by co-immunoprecipitation of cellular thioesterase and Nef stably expressed in CEM T lymphocytes. We examined by quantitative two-hybrid assay the interaction with thioesterase of Nef alleles known to be deficient in CD4-down-regulation. These Nef alleles were found partially or completely deficient for interaction with thioesterase whereas the Nef alleles which down-regulate CD4 still interact with thioesterase. We then selected by two-hybrid assay Nef point mutants unable to interact with thioesterase. These mutants were found to be completly deficient in CD4-down-regulation. These results suggest a correlation between CD4 down-regulation activity of Nef and its ability to interact with thioesterase.
Publication Types:
Keywords:
- ACOT8 protein, human
- Antigens, CD4
- Cloning, Molecular
- Coatomer Protein
- Down-Regulation
- Drug Interactions
- Fatty Acid Synthetase Complex
- Gene Products, nef
- HIV-1
- Humans
- In Vitro
- T-Lymphocytes
- Thiolester Hydrolases
- genetics
- immunology
- reverse transcriptase, Human immunodeficiency virus 1
- thioesterase II
Other ID:
UI: 102225347
From Meeting Abstracts