HIV-1 Tat down-regulates expression of mitotic centromere-associated kinesin through inhibiting promoter activity
10.3724/SP.J.1008.2010.00349
- Author:
Yue-Cheng HUANG
1
Author Information
1. Department of Radiation Medicine
- Publication Type:Journal Article
- Keywords:
MCAK;
Promoter activity;
Tat;
Te671 cell
- From:
Academic Journal of Second Military Medical University
2010;31(4):349-353
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the regulatory effect of HIV-1 Tat on mitotic centromere-associated kinesin (MCAK) expression and the related mechanism. Methods: Tat-expression TE671 cell model and purified prokaryotically expressed Tat protein were used to verify the down-regulated expression of MCAK using Northern blotting and Western blotting analysis. Various DNA fragments in the promoter region of the MCAK gene were amplified with PCR, linked to the luciferase reporter plasmid, and then transferred into TE671 cells. Luciferase activity analysis was performed to measure the promoter activity of various DNA fragments, so as to determine the active promoter region of MCAK gene. Moreover, HIV-1 Tat protein was co-incubated with TE671 cells, and the promoter activity was detected to analyze the modulating effect of Tat on promoter activity in vitro. Results: The results of Northern blotting and Western blotting analysis indicated that the mRNA and protein levels of MCAK were strongly decreased by Tat protein. The core region of MCAK promoter was located in -399∼+1 bp region, and the promoter activity was strikingly inhibited by Tat protein. Conclusion HIV-1 Tat has a marked inhibitory effect on MCAK expression, which might be related to the decreased promoter activity caused by Tat protein.
