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Identification of an amino acid–base contact in the GCN4–DNA complex by bromouracil-mediated photocrosslinking

Nature volume 359, pages 650–652 (1992)Cite this article

Abstract

THE bZIP DNA-binding proteins are characterized by a 50-amino-acid DNA binding and dimerization motif, consisting of a highly basic DNA-binding region ('b') followed by a leucine zipper dimerization region ('ZIP')1. The best characterized bZIP DNA-binding protein is GCN4, a yeast transcriptional activator2–6. GCN4 binds to a 9-base-pair two-fold-symmetric DNA site, 5'−A−4T−3G−2A−1C0T+1C+2A+3T+4−3' (refs 7–10). A detailed model known as the 'induced helical fork' model has been proposed for the structure of the GCN4–DNA complex4. Using a site-specific bromouracil-mediated photocrosslinking method, we show here that the alanine at position 238 of GCN4 contacts, or is close to, the thymine 5-methyl of A·T at position +3 of the DNA site in the GCN4–DNA complex. Our results strongly support the induced helical fork model4. Our site-specific bromouracil-mediated photocrosslinking method requires no prior information regarding the structure of the protein or the structure of the protein–DNA complex and should be generalizable to DNA-binding proteins that interact with the DNA major groove11,12

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  1. Richard H. Ebright: To whom correspondence should be addressed

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  1. Department of Chemistry and Waksman Institute, Rutgers University, New Brunswick, New Jersey, 08855, USA

    Erich E. Blatter, Yon W. Ebright & Richard H. Ebright

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Blatter, E., Ebright, Y. & Ebright, R. Identification of an amino acid–base contact in the GCN4–DNA complex by bromouracil-mediated photocrosslinking. Nature 359, 650–652 (1992). https://doi.org/10.1038/359650a0

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