(Adopted from Abstract) Aberration in transcription regulation, which requires regulatory proteins recognizing specific sequences of DNA, plays a central role in the development of many diseases including cancer. A novel DNA binding motif has been identified which governs sequence specific DNA recognition in six recently identified transcription factors and DNA binding proteins. Unlike most of the DNA binding domains known to date, this new motif binds to the minor groove of DNA rather than the major groove. The overall goal of this project is to characterize the molecular basis for the sequence specific DNA recognition of this novel motif and to delineate the conformational changes in DNA induced by binding of the protein. In this study, the focus is the DNA binding domain of Mrf-2 a developmentally regulated protein recognizing the modulator of the intermediate-early gene of human cytomegalovirus (HMCV). It is proposed to use state-of-the-art solution nuclear magnetic resonance (NMR) methods, in conjunction with site-directed mutagenesis, to investigate the molecular details of the protein - DNA recognition. This study should not only improve understanding of the structure-function relationship of the transcription factors, which utilize this motif for DNA recognition, but also increase knowledge of how proteins recognize specific sequences of DNA via minor groove binding. A better understanding of the biochemical machinery that regulates the transcription of genes will increase ability to ultimately manipulate gene transcription for the treatment of various diseases.
