Site-directed mutagenesis of cis-acting element,CHR,in human NFBD1 promoter region
- VernacularTitle:人NFBD1启动子区域顺式作用元件CHR的定点突变分析
- Author:
Jiang ZHU
;
Huan LAN
;
Suling HONG
;
Youquan BU
- Publication Type:Journal Article
- Keywords:
a nuclear factor with BRCT domain 1;
Promoter;
transcriptional regulation;
cell cycle genes homology region;
cis-acting element
- From:Journal of Third Military Medical University
2003;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of a cis-acting element,cell cycle genes homology region (CHR),in the transcriptional regulation of human NFBD1 gene (a nuclear factor with BRCT domain 1) by conducting a site-directed mutagenesis analysis on the element in human NFBD1 promoter region.Methods Wild type of NFBD1 promoter reporter,NFBD1-PS1-325,was used as template to make CHR mutant by using PCR based site-directed mutagenesis.Dual luciferase reporter assay was used to determine promoter reporter activity.Adiamycin treatment was employed to investigate the role of CHR in the transcriptional downregulation of NFBD1 after DNA damage.Results Site-directed mutagenesis of CHR caused a significant decrease in NFBD1 promoter activity,and also attenuated the transcriptional down-regulation of NFBD1 after DNA damage.Conclusion CHR element might be involved in both basic transcriptional regulation and transcriptional downregulation of NFBD1 after DNA damage.