Type

Text

Type

Thesis

Advisor

Luk, Ed | Hollingsworth, Nancy M

Date

2016-12-01

Keywords

Biochemistry -- Molecular biology

Department

Department of Biochemistry and Cell Biology

Language

en_US

Source

This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.

Identifier

http://hdl.handle.net/11401/76892

Publisher

The Graduate School, Stony Brook University: Stony Brook, NY.

Format

application/pdf

Abstract

The Kin28 subunit of the general transcription factor complex, TFIIH, is a kinase that phosphorylates the heptapeptide repeats at serine position 5 (Ser5) on the C-terminal domain of RNA polymerase II (Pol II). Optimal Ser5 phosphorylation is important during transcription for Pol II to transition from the initiation step to the elongation step in a process called promoter escape. During transcription initiation, the nucleosome directly downstream of the promoter, called +1, must be disassembled before Pol II can engage the transcription start site. Previous work showed that the transcription machinery plays a direct role in the disassembly of the +1 nucleosome, but the exact mechanism is currently unknown. We hypothesize that the disassembly of the +1 nucleosome occurs during promoter escape. To test this idea, I attempted to conditionally deplete Kin28, in the yeast, Saccharomyces cerevisiae, to block promoter escape. I then applied a modified genomic approach to map nascent Pol II-associated RNA molecules before and after Kin28 depletion. The Kin28 depletion failed to block promoter escape, possibly due to incomplete removal of Kin28 from the nucleus. Instead, I found that the Kin28 depleted strain accumulated Pol II-associated read-through transcripts beyond the transcription termination site suggesting that Kin28 plays a role in the processing of the nascent RNA of protein-coding genes. | 49 pages

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