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Cell cycle-regulated expression of mammalian CDC6 is dependent on E2F
G Hateboer1, A Wobst, B O Petersen
1Department of Experimental Oncology, European Institute of Oncology, 20141 Milan, Italy.
Molecular and Cellular Biology
|October 17, 1998
Summary
The E2F transcription factor regulates human CDC6 gene expression, crucial for DNA replication initiation. This finding links cell cycle control to DNA synthesis, highlighting E2F
Area of Science:
- Molecular Biology
- Cell Cycle Regulation
- Gene Expression
Background:
- E2F transcription factors regulate cell growth and proliferation genes in multicellular organisms.
- MBF and SBF complexes in yeast control cell cycle genes, with CDC6 as a target.
- Yeast Cdc6p and S. pombe Cdc18p homologs are essential for DNA replication initiation.
Purpose of the Study:
- To investigate the role of E2F in regulating the human CDC6 gene.
- To determine if human CDC6 is essential for DNA synthesis and cell cycle progression.
Main Methods:
- Cloning and analysis of the human CDC6 promoter.
- In vivo footprinting to identify E2F binding sites.
- Microinjection of anti-CDC6 antiserum and cotransfection experiments.
Main Results:
- Human CDC6 transcription is dependent on E2F, with E2F sites occupied in resting and growing cells.
- E2F downregulates and upregulates CDC6 expression during the cell cycle.
- Human CDC6 is essential and limiting for DNA synthesis; it cooperates with cyclin E for S-phase entry.
Conclusions:
- E2F directly regulates human CDC6 expression, linking G1 cell cycle control to DNA replication initiation.
- Human CDC6 is a critical limiting factor for DNA replication.
- The pRB pathway's regulation of cell cycle progression is directly connected to the expression of DNA replication proteins.