Abstract
Dbf4 is the regulatory subunit of Cdc7 kinase, which is essential for entry into and traversing through S phase. The level of Dbf4, which is critical for the activation of Cdc7, is regulated by transcription and protein degradation. To gain a better understanding as to how the transcription of human Dbf4 (HuDbf4) is regulated, we have cloned and characterized its promoter. We found that HuDbf4 core promoter is localized within −211 to −285 of the translation start-codon. This 75 bp DNA segment contains, among others, a putative MluI Cell-cycle Box (MCB). A point mutation within the MCB dramatically reduced the promoter activity. This is the first example that an MCB element plays an essential role in the activation of a core promoter in mammalian cells. The auxiliary elements required for the full promoter activity are present within 162-bp upstream from the core promoter (i.e., −286/−447). A point mutation within the Sp1 element at −353/−361 resulted in a decrease of promoter activity to the basal level, while the deletion of the putative HES-1 at −326/−331 dramatically increased the promoter activity. Taken together, our data suggests that the MCB element is essential for the core promoter activation, while the Sp1 positive regulator and the HES-1 repressor coordinately determine the efficiency of the HuDbf4 promoter. We have also found: (i) that the major transcription initiations occur at −220, −235 and −245; (ii) that HuDbf4 gene consists of 12 exons, which spread over a 33-kb region.
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Acknowledgements
We thank the members of the Lee lab, particularly, B Guo and A Pearce for many helpful discussions. This work was supported by the operating grants from the Canadian Institutes of Health Research (MT-15015), Northern Cancer Research Foundation, and the Northern Ontario Heritage Fund Corporation (NOHFC #50070). H Lee is a recipient of the Premier's Research Excellence Award.
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Wu, X., Lee, H. Human Dbf4/ASK promoter is activated through the Sp1 and MluI cell-cycle box (MCB) transcription elements. Oncogene 21, 7786–7796 (2002). https://doi.org/10.1038/sj.onc.1205914
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DOI: https://doi.org/10.1038/sj.onc.1205914