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Regulation Of Transcription By RNA Polymerase II In S. pombe

Gopalan, Sneha (2018). Regulation Of Transcription By RNA Polymerase II In S. pombe. PhD thesis. The Open University.

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An understanding of the mechanisms underlying the various stages of transcription is crucial to find solutions to the problems caused by mis-expression of genes that may give rise to a host of human diseases. My thesis research focusses on an analysis of the RNA Polymerase II elongation factor Ell1/Eaf1 in S. pombe. Eleven-nineteen lysine-rich in leukemia (ELL) is encoded by a gene involved in translocations with MLL in leukemia and forms a tight complex with ELL-associated factors (EAF). ELL/EAF is an RNA polymerase II elongation factor that in metazoa can assemble into a larger assembly that also includes P-TEFb and other proteins encoded by genes involved in MLL translocations. This larger assembly, sometimes called SEC, binds to a specific "docking site" in the metazoan Mediator complex. Distantly related ELL- and EAF-like genes were identified in S. pombe that encode Ell1/Eaf1 and can stimulate Pol II elongation in vitro. My thesis addresses two distinct projects, with overlapping motivations:

First, to see whether S. pombe might provide a good model for functional studies of the Mediator and ELL/EAF interaction, I carried out a thorough proteomic analysis of S. pombe Mediator and defined several new subunits. My results were recently published as part of a collaborative structural analysis of S. pombe Mediator.

Second, I used a combination of biochemical, genetic, and genomic approaches to characterize Ell1/Eaf1 function in fission yeast. Using mass spectrometry, I identified an uncharacterized sequence orphan, SPAC6G9.15c, that associates with both Ell1 and Eaf1 to form a ternary complex that, based on ChIP-seq localizes at genes with high RNA Pol II occupancy. I also performed an SGA screen for genes that genetically interact with ell1, eaf1, and SPAC6G9.15c and identified a set of overlapping genes that interact with all three, as well as others that interact only with ell1.

Item Type: Thesis (PhD)
Copyright Holders: 2017 The Author
Keywords: Schizosaccharomyces pombe; RNA polymerases; genetic transcription regulation; lysine
Academic Unit/School: Faculty of Science, Technology, Engineering and Mathematics (STEM)
Associated Research Centre: Stowers Institute for Medical Research
Item ID: 53696
Depositing User: Sneha Gopalan
Date Deposited: 30 Apr 2018 10:53
Last Modified: 18 Jan 2020 08:10
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