CCAAT-box binding protein NF-Y (CBF, CP1) recognizes the minor groove and distorts DNA.
The CCAAT box is one of the most common promoter elements. The evolutionarily 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 oligonucleotides 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, epsilon-globin and MSV, indicate that NF-Y is able to distort the double helix by angles of 62-82 degrees, depending on the site used, and suggest that nucleotides flanking the CCAAT pentanucleotide influence the degree of bending.
Dipartimento di Genetica e Biologia del Microganismi, Università di Milano, Italy.
A
A
Ronchi
M
M
Bellorini
N
N
Mongelli
R
R
Mantovani
D001483:Base Sequence; D001665:Binding Sites; D005075:Biological Evolution; D022762:CCAAT-Enhancer-Binding Proteins; D017124:Conserved Sequence; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009838:Oligodeoxyribonucleotides / Q000737:chemistry / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics; D015183:Restriction Mapping; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
D022762:CCAAT-Enhancer-Binding Proteins; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D011506:Proteins; D014157:Transcription Factors; D004247:DNA