Elucidating the mechanism of phenylpropanoid regulation by the Arabidopsis mediator complex
Abstract
The Mediator complex is a multi-protein co-regulator of eukaryotic transcription which plays a role in the expression of many, if not most, genes of the cell. Two Mediator subunits, REF4 and RFR1, were demonstrated to be important for the normal regulation of phenylpropanoid metabolism in Arabidopsis. Phenylpropanoids are a family of specialized plant metabolites derived from the amino acid phenylalanine and are involved in defense against pathogens, UV protection and structural support. In order to understand how Mediator regulates phenylpropanoid metabolism through REF4 and RFR1, a better basic understanding of this protein complex is required. Here I provide data on the initial steps in characterizing the Arabidopsis Mediator complex. First, I performed partial purification of Mediator using ion-exchange chromatography followed by LC-MS analysis of cation-exchange purified fraction. Then we generated and evaluated antibodies against specific Mediator subunits. Lastly, I used yeast two-hybrid to evaluate previously identified putative interactors of REF4, evaluated interaction between Mediator tail subunits with REF4 and RFR1, and identified new interaction candidates for RFR1.
Degree
M.S.
Advisors
Chapple, Purdue University.
Subject Area
Biochemistry
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