GENOME-WIDE DEPENDENCY OF TRANSCRIPTION MACHINERY ON GCN5 IN SACCHAROMYCES CEREVISIAE
Open Access
- Author:
- Cantor, David John
- Area of Honors:
- Biochemistry and Molecular Biology
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Benjamin Franklin Pugh, Thesis Supervisor
Benjamin Franklin Pugh, Thesis Supervisor
David Scott Gilmour, Thesis Honors Advisor
Dr. Wendy Hanna-Rose, Faculty Reader - Keywords:
- transcription machinery genome-wide mapping
Gcn5 deletion
ChIP-exo
SOLiD sequencing - Abstract:
- Eukaryotic gene regulation has largely been evolutionary conserved. The basic transcription machinery present in Saccharomyces cerevisiae, budding yeast, have close homologs in higher eukaryotic organisms. The Spt-Ada-Gcn5 (SAGA) complex is the major H3 histone acetyltransferase (HAT). While certain factor binding locations are known on a genomic scale, there exists little data on the functional dependencies of the transcriptional machinery during PIC assembly genome-wide. In the present study, genome-wide occupancies of 12 gene regulatory proteins in Gcn5 deletion strains were mapped using chromatin immunoprecipitation followed by next generation SOLiD sequencing (ChIP-seq) to better elucidate their dependencies on the HAT activity of SAGA. Gcn5 was deleted from strains with 12 gene regulatory proteins TAP-tagged. Stress state occupancy was compared to non-stress state occupancy by putting the cells through a heat shock. Evidence suggests that SAGA is involved in the stress response, and so a better elucidation of functional dependency on Gcn5 may be assessed upon heat shock stress. The following study focuses on analyzing the initial sequencing data from the deletion strains. We also show different genome-wide occupancy patterns for Rsc9, a subunit of RSC (Remodel the Structure of Chromatin) complex, and RNA polymerase II when at stress state compared to non-stress state in the Gcn5 deletion models suggesting a unique response to stress separate from the SAGA dependent pathway.