Regulation Of Transcription By RNA Polymerase II In <i>S. pombe</i>

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 <i>S. pombe</i>. 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 <i>S. pombe</i> that encode Ell1/Eaf1 and can stimulate Pol II elongation <i>in vitro</i>. My thesis addresses two distinct projects, with overlapping motivations: <br></br><br></br> First, to see whether <i>S. pombe</i> might provide a good model for functional studies of the Mediator and ELL/EAF interaction, I carried out a thorough proteomic analysis of <i>S. pombe</i> Mediator and defined several new subunits. My results were recently published as part of a collaborative structural analysis of <i>S. pombe</i> Mediator. <br></br><br></br> 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, <i>SPAC6G9.15c</i>, 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 <i>ell1, eaf1</i>, and <i>SPAC6G9.15c</i> and identified a set of overlapping genes that interact with all three, as well as others that interact only with <i>ell1</i>.

Name dc:type.qualificationname

phd

Level dc:type.qualificationlevel

doctoral

Grantor dc:publisher.institution

The Open University

Year dc:date.issued

2018

Author dc:creator

• Gopalan, Sneha

Language dc:language

en