Related Experiment Videos
The DNA-dependent ATPase activity associated with the class II basic transcription factor BTF2/TFIIH
R Roy1, L Schaeffer, S Humbert
1UPR 6520 (Centre National de la Recherche Scientifique), Unité 184 (Institut National de la Santé et de la Recherche Médicale), Faculté de Médecine, Strasbourg, France.
The Journal of Biological Chemistry
|April 1, 1994
Summary
The basal transcription factor BTF2/TFIIH contains a DNA-dependent ATPase and helicase activity, distinct from its associated carboxyl-terminal domain kinase. This ATPase and helicase function are linked to the p89/ERCC3 subunit.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Basal transcription factors like BTF2/TFIIH are essential for gene expression.
- These factors are often associated with enzymatic activities, including protein kinases.
Purpose of the Study:
- To investigate the enzymatic activities associated with the human basal transcription factor BTF2/TFIIH.
- To determine the relationship between the DNA-dependent ATPase, helicase, and carboxyl-terminal domain kinase activities within BTF2/TFIIH.
Main Methods:
- Biochemical assays to characterize DNA-dependent ATPase and helicase activities.
- Inhibition studies using Sarkosyl and staurosporine.
- Nucleotide competition experiments.
- Analysis of recombinant wild-type and mutated p89/ERCC3 polypeptides.
Main Results:
- BTF2/TFIIH possesses both DNA-dependent ATPase and helicase activities, localized to the p89 subunit.
- These ATPase/helicase activities are biochemically distinct from the associated carboxyl-terminal domain kinase.
- The ATPase and helicase functions are demonstrated to be interconnected.
Conclusions:
- The p89/ERCC3 subunit of BTF2/TFIIH harbors coupled ATPase and helicase activities crucial for transcription.
- BTF2/TFIIH integrates DNA unwinding and kinase functions for RNA polymerase II regulation.