Cell Cycle-dependent Switch of Up- and Down-regulation of Human hsp70 Gene Expression by Interaction between c-Myc and CBF/NF-Y*

A CCAAT box-binding protein subunit, CBF-C/NF-YC, was cloned as a protein involved in the c-Myc complex formed on the G1-specific enhancer in the human hsp70gene. CBF-C/NF-YC directly bound to c-Myc in vitro andin vivo in cultured cells. The CBF/NF-Y·c-Myc complex required the HSP-MYC-B element as well as CCAAT in the hsp70G1-enhancer, while the purified CBF subunits recognized only CCAAT even in the presence of c-Myc. Both the HSP-MYC-B and CCAAT elements were also required for the enhancer activity. In transient transfection experiments, the CBF/NF-Y·c-Myc complex, as well as transcription due to the G1-enhancer, was increased by the introduction of c-Myc at low doses but decreased at high doses. The repression of both complex formation and transcription by c-Myc at high doses was abrogated by the introduction of CBF/NF-Y in a dose-dependent manner. Furthermore, the CBF/NF-Y·c-Myc complex bound to the G1-enhancer appeared in the early G1 phase of the cell cycle when c-Myc was not higly expressed and gradually disappeared after the c-Myc expression reached its maximum. The results indicate that the cell cycle-dependent expression of the hsp70 gene is regulated by the intracellular amount of c-Myc through the complex formation states between CBF/NF-Y and c-Myc.

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