摘要
DNA polymerase delta is encoded by the POLD1 gene,the transcription of which is strictly cell cycle-dependent.However,the means by which POLD1 transcription is regulated by the cell cycle mechanism is currently unknown.We discovered a novel element in the POLD1 promoter known as a CDE(cell cycle-dependent element)/CHR(cell cycle gene homology region) element.A series of luciferase reporter constructs containing various POLD1 promoter mutations were used to investigate the role of the CDE/CHR element in POLD1 transcription.When the CDE/CHR element was mutated,the promoter activity was up-regulated,and the cell-cycle related factors E2F1 and p21 stopped regulating the promoter.Furthermore,cell cycle-dependent changes in the promoter activity required the integrative CDE/CHR element.Electrophoretic mobility shift assay(EMSA) revealed the presence of at least three types of DNA/protein complexes binding to the CDE/CHR element.Our findings provide strong evidence that the CDE/CHR-like sequence is an active functional element in the POLD1 promoter,which is important for the cell cycle regulation of the POLD1 gene.
DNA polymerase delta is encoded by the POLD1 gene,the transcription of which is strictly cell cycle-dependent.However,the means by which POLD1 transcription is regulated by the cell cycle mechanism is currently unknown.We discovered a novel element in the POLD1 promoter known as a CDE(cell cycle-dependent element)/CHR(cell cycle gene homology region) element.A series of luciferase reporter constructs containing various POLD1 promoter mutations were used to investigate the role of the CDE/CHR element in POLD1 transcription.When the CDE/CHR element was mutated,the promoter activity was up-regulated,and the cell-cycle related factors E2F1 and p21 stopped regulating the promoter.Furthermore,cell cycle-dependent changes in the promoter activity required the integrative CDE/CHR element.Electrophoretic mobility shift assay(EMSA) revealed the presence of at least three types of DNA/protein complexes binding to the CDE/CHR element.Our findings provide strong evidence that the CDE/CHR-like sequence is an active functional element in the POLD1 promoter,which is important for the cell cycle regulation of the POLD1 gene.
作者
SONG NanMeng1,ZHU XiaoYu1,2,SHI Lei1,AN Jing1,WU YanWei1 & SANG JianLi1 1 Key Laboratory of Cell Proliferation and Regulation of Ministry of Education,Beijing Normal University,Beijing 100875,China
2 State Key Laboratory for Infectious Disease Prevention and Control,Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China
基金
Supported by National Basic Research Program of China (Grant No. 2007CB914401)
National High Technology Research and Development Program of China (Grant No. 2006AA02Z4A6)
