Structure and promoter activity of the mouse CDC25A gene.
CDC25A is a member of a group of highly related, dual-specificity phosphatases that promote cell cycle phase transitions by regulating the activity of cyclin-dependent kinases. Here we report the cloning and genomic sequence of 21,067 nucleotides encompassing the mouse CDC25A gene. The coding sequence is expressed from 17,904 bp of genomic DNA comprising 15 exons. We also mapped the transcription initiation site to a consensus initiator element proximal to an SP1 site. Approximately 1 kb of sequence upstream of the transcription initiation site confers promoter activity and cell type specificity to a reporter gene construct. Surprisingly, transcription from this promoter was repressed by over-expression of catalytically active but not catalytically inactive CDC25A protein. We also show, using NIH 3T3 cells, that murine CDC25A mRNA levels fluctuate only modestly over the cell cycle. Our findings provide insights into the regulation of CDC25A expression and have facilitated construction of gene knock-out vectors.
Paskind
M
M
BASF Bioresearch Corp, Worcester, Massachusetts 01605-4314, USA.
Johnston
C
C
Epstein
P M
PM
Timm
J
J
Wickramasinghe
D
D
Belanger
E
E
Rodman
L
L
Magada
D
D
Voss
J
J
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle / Q000502:physiology; D002460:Cell Line; D018076:DNA, Complementary; D005786:Gene Expression Regulation / Q000502:physiology; D018390:Gene Targeting; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014158:Transcription, Genetic; D020687:cdc25 Phosphatases / Q000737:chemistry / Q000235:genetics*
D018076:DNA, Complementary; C517004:CDC25A protein, human; C517005:Cdc25a protein, mouse; D020687:cdc25 Phosphatases