Mapping of DNA-binding proteins along the yeast genome by UV-induced DNA-protein crosslinking.

UV-induced crosslinking of DNA-binding proteins to DNA in intact nuclei of Saccharomyces cerevisiae and subsequent 'protein image' hybridization were applied to map non-histone proteins along single-copy genes of yeast. We detected two polypeptides that most probably correspond to core subunits of yeast RNA-polymerase II in the coding region of transketolase gene (TKL2). Several non-histone proteins were also detected which bind to the upstream region of TKL2 gene, and to the intergenic spacer between calmodulin (CMD1) and beta-mannosyl transferase (ALG1) genes.

[Detection of nonhistone proteins, bound with regulatory elements of yeast rRNA genes, by UV-crosslinks in intact nuclei].

We have studied the arrangement of DNA-binding proteins along yeast ribosomal non-transcribed spacer by UV-induced DNA-protein crosslinking on intact nuclei. We show binding of proteins with apparent Mw 120 and 30 kDa in promoter region, 120 kDa in enhancer region, 40 kDa in ARS. These proteins were identified preliminary as REB1 (promoter, enhancer), TFID (promoter), MCM3 (ARS).