Characterization of the Enhancer and Promoter Elements of the Human TAF(II)55 Gene

To understand the mechanisms by which activators and cofactors function on TATA-less promoters, we also purified a transcriptional cofactor complex, Mediator, by first establishing a HeLa-derived cell line (hMED7-7) that expresses the epitope-tagged Med7 component of human Mediator. From the derived nuclear extracts, or the 500 mM and the 850 mM KCl P11 phosphocellulose column fractions of the derived nuclear extracts, three forms of Mediator complexes containing previously characterized Mediator subunits were purified. The 500 mM fraction-derived Mediator contains Cdk8/Cyclin C, while the 850 mM fraction-derived Mediator lacks the protein pair and likely corresponds to the previously characterized positive cofactor 2 (PC2). The associated RNA polymerase II was also depleted from the Mediator complexes with an immobilized antibody against the largest subunit of RNA polymerase II. The availability of these human Mediator complexes allows us to address the functional redundancy and/or transcriptional synergy among three general cofactors (TAFIIs, Mediator and PC4) commonly implicated in activator-dependent transcription from both TATA-containing and TATA-less promoters.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biochemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2015

Author dc:creator

• Zhou, Tianyuan

Contributors dc:contributor

• Cheng-Ming Chiang

• Biology, Molecular

Language dc:language

eng

Identifier

(MiAaPQ)AAI3044274

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/84782