Selective proteolysis defines two DNA binding domains in yeast transcription factor tau

Abstract

Transcription of eukaryotic transfer RNA genes involves, as a primary event, the stable binding of a protein factor to the intragenic promoter. The internal control region is composed of two non-contiguous conserved sequence elements, the A and B blocks. These are variably spaced depending on the genes. tau, a large transcription factor purified from yeast cells, interacts with these two control elements as shown by DNase I footprinting, exonuclease digestion, dimethyl sulphate protection experiments and by analysis of point mutations. Here we used a limited proteolysis treatment to obtain a smaller form of tau with drastically altered DNA binding properties. A protease-resistant domain interacts solely with the B block region of tRNA genes.