Regulated expression of HIV-1 Rev function in mammalian cell lines
作者:
Lauren Swenarchuk,
Penelope Harakidas,
Alan Cochrane,
期刊:
Canadian Journal of Microbiology
(NRC Available online 1999)
卷期:
Volume 45,
issue 6
页码: 480-490
ISSN:0008-4166
年代: 1999
DOI:10.1139/w99-011
出版商: NRC Research Press
数据来源: NRC
摘要:
In order to facilitate further investigation of Rev function, we have generated two systems for the inducible expression of Rev in mammalian cell lines (HeLa and U937) using either a tetracycline-regulated promoter or fusion of Rev to a modified form of the hormone binding domain of the estrogen receptor. In the case of the fusion of Rev to the modified hormone binding domain of the estrogen receptor, we demonstrated induction of Rev function in response to tamoxifen administration to levels comparable to that of the unmodified Rev protein. Subsequently, U937 lines were generated that retained the observed pattern of hormone-dependent function of the Rev fusion protein. In the case of the tetracycline-regulated system, cell lines (both HeLa and U937) were generated that displayed tight regulation of Rev. In the case of the HeLa cell lines, they were used for the subsequent generation of stable cell lines expressing either HIV-1envor chloramphenicol acetyl transferase (CAT) in a Rev-dependent fashion. Using the latter cell lines, we demonstrate the ability to control Rev expression over a broad concentration range and find that, as soon as Rev expression is detectable, induction of Rev-dependent gene expression is also observed.Key words: Rev, tamoxifen, tetracyline-regulated transcription, Rev function, threshold effects.
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