Identification of sequence-specific DNA-binding factors by label transfer: application to the adenovirus-2 major late promoter | Zendy

Л. Г. Николаев | Zendy; Boris O. Glotov | Zendy; A. V. Belyavsky | Zendy; S. A. Grachev | Zendy; A. V. Levin | Zendy

Open Access

Identification of sequence-specific DNA-binding factors by label transfer: application to the adenovirus-2 major late promoter

Author(s) -

Л. Г. Николаев,

Boris O. Glotov,

A. V. Belyavsky,

S. A. Grachev,

A. V. Levin

Publication year - 1988

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/16.2.519

Subject(s) - biology , dna , identification (biology) , computational biology , sequence (biology) , dna binding site , adenoviridae , base sequence , genetics , dna binding protein , dna sequencing , microbiology and biotechnology , promoter , transcription factor , gene , recombinant dna , gene expression , botany

A method of affinity labelling proteins specifically associated with DNA target sequences is proposed. The method utilizes covalent UV-crosslinking of proteins to highly labelled DNA (e.g. in crude cell or nuclear extracts) followed by degradation of the DNA to short oligonucleotides. Proteins selectively labelled by attached residual oligonucleotides are readily amenable to molecular mass determination. Using this approach, we have characterized a HeLa polypeptide specifically bound to a short segment of the adenovirus-2 major late promoter (Ad2 MLP). A molecular mass value (approximately 51 kD) and precise location of the crosslinking site(s) of the protein within the MLP (-55 with respect to the cap site) were determined.

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