The DNA-dependent ATPase activity associated with the class II basic transcription factor BTF2/TFIIH.
BTF2/TFIIH from human, delta from rat, and factor b from yeast are multisubunit basal transcription factors that have been shown to be closely associated with a protein kinase capable of phosphorylating the carboxyl-terminal domain of the large subunit of RNA polymerase II (Lu, H., Zawel, L., Fischer, L., Egly, J. M., and Reinberg, D. (1992) Nature 358, 641-645; Serizawa, H., Conaway, R. C., and Conaway, J. W. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 7476-7480; Feaver, W. J., Gileadi, O., and Kornberg, R. D. (1991) Cell 67, 1223-1230). We report here that a DNA-dependent ATPase and the previously characterized helicase (Schaeffer, L., Roy, R., Humbert, S., Moncollin, V., Vermeulen, W., Hoeijmakers, J., Chambon, P., and Egly, J. M. (1993) Science 260, 58-63) are both associated with BTF2 and reside with the p89 polypeptide subunit. The DNA requirement, the effect of Sarkosyl and staurosporine inhibitors, as well as nucleotide competition experiments, clearly distinguished ATPase/helicase from the carboxyl-terminal domain kinase. Using recombinant wild type or mutated p89/ERCC3 polypeptides and different forms of DNA template, we show the connection between ATPase and the helicase.
Roy
R
R
UPR 6520 (Centre National de la Recherche Scientifique), Unité 184 (Institut National de la Santé et de la Recherche Médicale), Faculté de Médecine, Strasbourg, France.
Schaeffer
L
L
Humbert
S
S
Vermeulen
W
W
Weeda
G
G
Egly
J M
JM
D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism*; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D004265:DNA Helicases / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D012521:Sarcosine / Q000031:analogs & derivatives / Q000494:pharmacology; D019311:Staurosporine; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism*; D019473:Transcription Factors, TFII
D000470:Alkaloids; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C490039:TAF6 protein, S cerevisiae; D035381:TATA-Binding Protein Associated Factors; D035362:Transcription Factor TFIID; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D051758:Transcription Factor TFIIH; C025231:sarkosyl; D011494:Protein Kinases; C060137:carboxy-terminal domain kinase; D011493:Protein Kinase C; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D019311:Staurosporine; D012521:Sarcosine