Incorporation of 5-bromodeoxycytidine in the adenovirus 2 replication origin interferes with nuclear factor 1 binding.
We have studied the binding of nuclear factor 1 (NFI), a human sequence-specific DNA-binding protein, to a DNA fragment substituted in vitro with 5-bromodeoxycytidine (5-BrdC). Even at low substitution grades binding of NFI to its recognition sequence was considerably lower than with the unsubstituted control fragment. We 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 in 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.
de Vries
E
E
Laboratory for Physiological Chemistry, State University of Utrecht, The Netherlands.
Bloemers
S M
SM
van der Vliet
P C
PC
D000256:Adenoviridae / Q000235:genetics*; D001972:Bromodeoxycytidine / Q000494:pharmacology*; D022762:CCAAT-Enhancer-Binding Proteins; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003841:Deoxycytidine / Q000031:analogs & derivatives*; D005809:Genes, Regulator; D005814:Genes, Viral; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D011485:Protein Binding / Q000187:drug effects; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1
D022762:CCAAT-Enhancer-Binding Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D051839:NFI Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D051840:Y-Box-Binding Protein 1; C070856:YBX1 protein, human; D003841:Deoxycytidine; D001972:Bromodeoxycytidine