Regulation of gene expression is fundamental to living processes such as growth control and development, and improved knowledge of the general transcription machinery is essential to understanding gene regulation. We have identified an RNA polymerase II holoenzyme in yeast which appears to be the form of RNA polymerase II that initiates transcription in vivo. This holoenzyme contains RNA polymerase II, general transcription factors, and a multiprotein complex composed of SRB, SWI and SNF proteins. The SRB/SWI/SNF complex is associated with the RNA polymerase II large subunit C-terminal domain (CTD) and has roles in the response to transcriptional activators and repressors. A similar holoenzyme was recently identified in mammalian cells. The experiments described in this proposal are designed to investigate the roles of RNA polymerase II holoenzyme components in control of class II gene expression using a combination of genetic and biochemical approaches in yeast. To accomplish this goal, the four specific aims of this proposal are: 1) to identify all of the CTD- associated components of the yeast RNA polymerase II holoenzyme; 2) to survey the yeast genome to identify the set of promoters that require SRB/SWI/SNF function in vivo; 3) to investigate further the subunit structure and function of the SRB/SWI/SNF subcomplex, and 4) to investigate how positive and negative regulators interact with the regulate SRB/SWI/SNF components of the holoenzyme. The health relatedness of this project derives from its contribution to the understanding of basic molecular mechanisms that control gene expression.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034365-15
Application #
2838505
Study Section
Genetics Study Section (GEN)
Program Officer
Tompkins, Laurie
Project Start
1984-12-01
Project End
2000-11-30
Budget Start
1998-12-01
Budget End
1999-11-30
Support Year
15
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Whitehead Institute for Biomedical Research
Department
Type
DUNS #
076580745
City
Cambridge
State
MA
Country
United States
Zip Code
02142
Ng, Huck Hui; Robert, Francois; Young, Richard A et al. (2003) Targeted recruitment of Set1 histone methylase by elongating Pol II provides a localized mark and memory of recent transcriptional activity. Mol Cell 11:709-19
Wyrick, J J; Aparicio, J G; Chen, T et al. (2001) Genome-wide distribution of ORC and MCM proteins in S. cerevisiae: high-resolution mapping of replication origins. Science 294:2357-60
Geisberg, J V; Holstege, F C; Young, R A et al. (2001) Yeast NC2 associates with the RNA polymerase II preinitiation complex and selectively affects transcription in vivo. Mol Cell Biol 21:2736-42
Robertson, L S; Causton, H C; Young, R A et al. (2000) The yeast A kinases differentially regulate iron uptake and respiratory function. Proc Natl Acad Sci U S A 97:5984-8
Kimmelman, J; Kaldis, P; Hengartner, C J et al. (1999) Activating phosphorylation of the Kin28p subunit of yeast TFIIH by Cak1p. Mol Cell Biol 19:4774-87
Hengartner, C J; Myer, V E; Liao, S M et al. (1998) Temporal regulation of RNA polymerase II by Srb10 and Kin28 cyclin-dependent kinases. Mol Cell 2:43-53
Woychik, N A; McKune, K; Lane, W S et al. (1993) Yeast RNA polymerase II subunit RPB11 is related to a subunit shared by RNA polymerase I and III. Gene Expr 3:77-82
McKune, K; Richards, K L; Edwards, A M et al. (1993) RPB7, one of two dissociable subunits of yeast RNA polymerase II, is essential for cell viability. Yeast 9:295-9
Choder, M; Young, R A (1993) A portion of RNA polymerase II molecules has a component essential for stress responses and stress survival. Mol Cell Biol 13:6984-91
Woychik, N A; Young, R A (1992) Genes encoding transcription factor IIIA and the RNA polymerase common subunit RPB6 are divergently transcribed in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 89:3999-4003

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