TATA Binding Protein Discriminates between Different Lesions on DNA, Resulting in a Transcription Decrease

ABSTRACT

DNA damage recognition by basal transcription factors follows different mechanisms. Using transcription-competition, nitrocellulose filter binding, and DNase I footprinting assays, we show that, although the general transcription factor TFIIH is able to target any kind of lesion which can be repaired by the nucleotide excision repair pathway, TATA binding protein (TBP)-TFIID is more selective in damage recognition. Only genotoxic agents which are able to induce kinked DNA structures similar to the one for the TATA box in its TBP complex are recognized. Indeed, DNase I footprinting patterns reveal that TBP protects equally 4 nucleotides upstream and 6 nucleotides downstream from the A-T (at position −29 of the noncoding strand) of the adenovirus major late promoter and from the G-G of a cisplatin-induced 1,2-d(GpG) cross-link. Together, our results may partially explain differences in transcription inhibition rates following DNA damage.

ACKNOWLEDGMENTS

We thank F. Jeffrey Dilworth, D. Moras, and P. Hanawalt for very fruitful discussions. We thank I. Kuraoka and R. Wood for supplying the DNA with a platinum adduct at a specific site and critical reading of the manuscript and A. Fery for her excellent technical assistance.

This work was supported by grants from the Human Frontier Program, the INSERM, the CNRS, the Hôpital Universitaire de Strasbourg, the Ministère de la Recherche et de l’Enseignement Supérieur, the Association pour la Recherche sur le Cancer, and La Ligue Nationale contre le Cancer.

F.C. and P.F. contributed equally to this work.