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 unknow...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.展开更多
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)
文摘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.
