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Nucleic Acids Research, 1995, Vol. 23, No. 22 4565-4572
© 1995

Articles

CCAAT-box binding protein NF-Y (CBF, CP1)recognizes the minor groove and distorts DNA

Antonella Ronchi, Marianna Bellorini, Nicola Mongelli1 and Roberto Mantovani*

Dipartimento di Genetica e Biologia dei Microrganismi, Universita di Milano Via Celoria 26, 20133 Milano, Italy 1Pharmacia/Farmitalia Carlo Erba, Nerviano Research Center Via Papa Giovanni XXIII 68, 20014 Nerviano, Italy

*To whom correspondence should be addressed

Received September 21, 1995. Accepted October 11, 1995.

The CCAAT box is one of the most common promoter elements. The evolutionary conserved heteromeric factor NF-Y binds this sequence with high affinity and specificity. By comparing the methylation interference patterns of different sites, performing electrophoretic mobility shift assays (EMSA) with IC-substituted ollgonucleotldes and competition experiments with the minor groove binding (MGB) drugs distamicin A, tallimustine and Hoechst 33258 we show that NF-Y makes key minor groove interactions. Circular permutation assays on four CCAAT boxes, MHC Class II Ea, HSP70, {varepsilon}-globin and MSV, indicate that NF-Y is able to distort the double helix by angles of 62-82° depending on the site used, and suggest that nucleotides flanking the CCAAT pentanucleotlde influence the degree of bending.


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