Cloning and characterization of Chinese hamster CDC7 (ChCDC7)

The Cdc7 serine/threonine kinase is essential for entry into and to traverse through S phase. We have cloned the putative Chinese hamster CDC7 (ChCDC7) cDNA that is capable of encoding a protein of 572 amino acids with predicted molecular mass of 62.6 kDa. The ChCdc7 protein includes all 11 kinase domains that are conserved among the Cdc7-related protein kinases. In addition, the ChCdc7 protein kinase contains at least two kinase inserts that show substantial identity to those of huCdc7p. Overall, ChCdc7p shares 81, 56, 30, and 27% amino acid sequence identity with the Cdc7-related proteins of human, Xenopus, Saccharomyces cerevisiae, and Schizosaccharomyces pombe, respectively. Although the levels of ChCDC7 mRNA and protein are relatively constant throughout the cell cycle in the cycling cells, they are extremely low in the cells synchronized in the quiescent stage (i.e., G0). When cells in G0 are released into the cell cycle, the levels of ChCDC7 mRNA and protein increase slowly until the cells reach the G1/S border, at which time the increase is rapid. This suggests that a number of signal transduction pathways may have to be activated prior to CDC7 gene expression. Interestingly, the ChCdc7-GFP fusion protein formed discrete granules in the nuclei of cells arrested in early S phase by aphidicolin, raising the possibility that ChCdc7p is part of the "replication factory."