This project addresses the intriguing question of how eukaryotic RNA polymerases transcribe through chromatin. The project involves development of the minimal in vitro system for transcription through nucleosome by Pol II, and Pol III from S. cerevisiae. RNAP from E. coli, which behaves very similarly to the eukaryotic enzyme in nucleosomal transcription, is used in this project as an easy-handled prototype of Pol II (Kireeva et al., Molecular Cell, 2002).

Agency
National Institute of Health (NIH)
Institute
Division of Basic Sciences - NCI (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC010343-04
Application #
6951653
Study Section
(GRCB)
Project Start
Project End
Budget Start
Budget End
Support Year
4
Fiscal Year
2003
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Kireeva, Maria L; Hancock, Brynne; Cremona, Gina H et al. (2005) Nature of the nucleosomal barrier to RNA polymerase II. Mol Cell 18:97-108
Studitsky, Vasily M; Walter, Wendy; Kireeva, Maria et al. (2004) Chromatin remodeling by RNA polymerases. Trends Biochem Sci 29:127-35
Walter, W; Kireeva, M L; Tchernajenko, V et al. (2003) Assay of the fate of the nucleosome during transcription by RNA polymerase II. Methods Enzymol 371:564-77
Walter, Wendy; Kireeva, Maria L; Studitsky, Vasily M et al. (2003) Bacterial polymerase and yeast polymerase II use similar mechanisms for transcription through nucleosomes. J Biol Chem 278:36148-56
Kireeva, Maria L; Lubkowska, Lucyna; Komissarova, Natalia et al. (2003) Assays and affinity purification of biotinylated and nonbiotinylated forms of double-tagged core RNA polymerase II from Saccharomyces cerevisiae. Methods Enzymol 370:138-55