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Nucleic Acids Research, 1987, Vol. 15, No. 18 7223-7234
© 1987

Articles

Incorporation of 5-bromodeoxycytidine in the adenovirus 2 replication origin interferes with nuclear factor 1 binding

Erik de Vries, S. Margreet Bloemers and Peter C. van der Vliet

Laboratory for Physiological Chemistry, State University of Utrecht Vondellaan 24a, 3521 GG Utrecht, The Netherlands

Received August 4, 1987. Accepted September 2, 1987.

We have studied the binding of nuclear factor 1 (NFI), a human sequence-specific DNA-b1nd1ng protein, to a DNA fragment substituted in vitro with 5-bromodeoxycytid1ne (5-BrdC). Even at low substitution grades binding of NFI to its recognition sequence was considerably lower than with the unsubstituted control fragment. He developed a procedure to cleave substituted DNA specifically at a BrdC residue and searched for contacts between NFI and 5-BrdC residues by an interference assay. Surprisingly, no specific contacts were found 1n or near the recognition sequence. It appeared instead that interference was inversely related to the distance of a 5-BrdC residue from the NFI binding site. Models to explain these results, including a possible sliding mechanism, are discussed.


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