IDENTIFICATION OF AN AMINO ACID-BASE CONTACT IN THE GCN4-DNA COMPLEX BY BROMOURACIL-MEDIATED PHOTO-CROSS-LINKING

被引:79
作者
BLATTER, EE
EBRIGHT, YW
EBRIGHT, RH
机构
[1] RUTGERS STATE UNIV,DEPT CHEM,NEW BRUNSWICK,NJ 08855
[2] RUTGERS STATE UNIV,WAKSMAN INST,NEW BRUNSWICK,NJ 08855
关键词
D O I
10.1038/359650a0
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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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页码:650 / 652
页数:3
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