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CBP/cycA, a CCAAT-binding Protein Necessary for Adhesion-dependent Cyclin A Transcription, Consists of NF-Y and a Novel M_r 115,000 Subunit¹

III. Medizinische Klinik, Klinikum Mannheim der Universität Heidelberg, 68305 Mannheim Germany [A. K.]; Department of Human Oncology, University of Wisconsin, Madison, Wisconsin 53792 [C-P. C.]; and McArdle Laboratory for Cancer Research, University of Wisconsin, Madison, Wisconsin 53706 [W. W. W., W. E. F.]

Cells of most tissues, with the exception of hematopoietic cells, require adhesion to an appropriate surface to grow. Cyclin A is needed for cell cycle progression at the G₁-S transition, and appearance of cyclin A mRNA and protein in late G₁ has been shown to be dependent on adhesion-initiated signals in normal rat kidney fibroblasts. Previously, we have reported that the adhesion-dependent activation of cyclin A transcription in late G₁ is mediated by CBP/cycA (CCAAT-binding protein for cyclin A gene), a novel CCAAT-binding protein. Specific binding of CBP/cycA, a M_r 30,000/40,000/115,000 heterotrimeric protein complex, to the CCAAT element of the cyclin A promoter was detectable in growing but not in G₀-arrested or nonadherent normal rat kidney cells. Here, we demonstrate that the M_r 30,000/40,000 subunits of CBP/cycA are identical with NF-Y_A and NF-Y_B, the two subunits of NF-Y. In addition, we show that, aside from CBP/cycA, NF-Y itself also binds to the CCAAT element of the cyclin A promoter. But, whereas the binding of CBP/cycA is adhesion and cell cycle dependent and correlates with the expression of cyclin A in late G₁ phase, NF-Y itself seems to bind in a cell cycle-independent manner.

¹ This work was supported by NIH Grant R37-CA42024. A. K. is the recipient of a postdoctoral fellowship from the Deutsche Krebshilfe.

² To whom requests for reprints should be addressed, at McArdle Laboratory for Cancer Research, University of Wisconsin, 1400 University Avenue, Madison, WI 53706. Phone: (608) 262-1275; Fax: (608) 262-2824; E-mail: fahl@oncology.wisc.edu.

Received 5/ 5/97. Accepted 9/12/97.

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