The regulation of eukaryotic genes involves many hundreds of proteins. How they work together at the many thousands of genes that comprise a genome is not known. Little is known about how this interplay changes as cellular genomes are reprogrammed in response to signaling events such as environmental stress. The work proposed here is a continuation of a project to broadly survey where transcriptional regulatory proteins are located throughout the genome of the model eukaryotic organism Saccharomcyes cerevisiae, and where they translocate to when the genome is reprogrammed by environmental signals, such as heat shock and other stresses. Heat shock provides a rapid and simple programming event for the cell. Current studies on this project have already revealed novel insights into gene regulation by demonstrating that pre-initiation complexes partially assemble in nucleosome-free promoter regions. However, full assembly requires removal of the "-1" nucleosome located on the upstream side of the promoter. The genomic location of ~200 gene- and chromatin-regulatory proteins have been determined at low resolution by ChIP-chip, under normal and heat shock conditions. The data is now being analyzed. High resolution mapping will be achieved by retrofitting the ChIP-chip libraries for ChIP-seq. Even higher resolution mapping will be achieved on a select set of factors using reagents to trim the ChIP DNA. Relationships among binding events will provide new insights into transcription complex assembly and regulation.

Public Health Relevance

Our cells are constantly faced with environmental extremes, involving temperature, starvation, radiation and harmful chemicals. How we deal with this stress depends upon the action of our transcription machinery on genes. Therefore a broad understanding of how our transcription machinery works in the face of various stresses is essential for a physiological understanding of human health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Environmental Health Sciences (NIEHS)
Type
Research Project (R01)
Project #
5R01ES013768-08
Application #
8599771
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Tyson, Frederick L
Project Start
2005-04-01
Project End
2014-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
8
Fiscal Year
2014
Total Cost
$295,385
Indirect Cost
$92,885
Name
Pennsylvania State University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
003403953
City
University Park
State
PA
Country
United States
Zip Code
16802
Chang, Gue Su; Chen, Xiangyun Amy; Park, Bongsoo et al. (2014) A comprehensive and high-resolution genome-wide response of p53 to stress. Cell Rep 8:514-27
Ghosh, Sujana; Pugh, B Franklin (2011) Sequential recruitment of SAGA and TFIID in a genomic response to DNA damage in Saccharomyces cerevisiae. Mol Cell Biol 31:190-202
Rhee, Ho Sung; Pugh, B Franklin (2011) Comprehensive genome-wide protein-DNA interactions detected at single-nucleotide resolution. Cell 147:1408-19
Venters, Bryan J; Wachi, Shinichiro; Mavrich, Travis N et al. (2011) A comprehensive genomic binding map of gene and chromatin regulatory proteins in Saccharomyces. Mol Cell 41:480-92
Samorodnitsky, Eric; Pugh, B Franklin (2010) Genome-wide modeling of transcription preinitiation complex disassembly mechanisms using ChIP-chip data. PLoS Comput Biol 6:e1000733
Venters, Bryan J; Pugh, B Franklin (2009) A canonical promoter organization of the transcription machinery and its regulators in the Saccharomyces genome. Genome Res 19:360-71
Venters, Bryan J; Pugh, B Franklin (2009) How eukaryotic genes are transcribed. Crit Rev Biochem Mol Biol 44:117-41