Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.590950
Title: The amino-terminal transactivation domain of the human androgen receptor : protein-protein interactions and structural characteristics
Author: Lavery, Derek Norman
Awarding Body: University of Aberdeen
Current Institution: University of Aberdeen
Date of Award: 2007
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Abstract:
An important target protein for the AR-AF1 domain is Transcription Factor IIF (TFIIF). At initiation of transcription, TFIIF recruits additional basal transcription factors, stabilises the transcriptional complex and increases elongation efficiency. Using chromatin immunoprecipitation, it was observed that the AR occupied distinct regions of the prostate-specificantigen enhancer but did not migrate with the elongating transcriptional complex. The major subunit of TFAAF, RAP74, has previously been shown to interact with AR-AF1 by our laboratory and it was observed that AR-AF1 can interact with both terminal regions of RAP74. Now, by selectively disrupting helices that structure the globular RAP74 C-terminal domain it appears that AR-AF1 binds to a groove within this region and specific hydrophobic amino acids are important in this generation. The kinetics of RAP74/AR-AF1 interactions have not been determined using surface plasmon resonance. Interestingly, AR-AF1 interacts differently with N- and C-terminal regions of RAP74 and the overall affinities are in the nanomolar range. The structural properties of AR-AF1 were examined using both fluorescence spectroscopy and gel filtration chromatography. It was found that the transactivation domain is structurally flexible and exists in a conformation that is not random coil or globular suggesting that it may be a molten globule. AR-AF1 interacted weakly with 8-anilinonaphthalene-1-sulfonic acid, a hydrophobic probe used to characterise the molten globule folding state. Gel filtration chromatography indicates that AR-AF1 is ∼65 kDa and has a hydrodynamic radius of ∼36 Co much larger than predictions suggest. Surprisingly, by plotting these properties on "folding curves", AR-AF1 is positioned alongside molten globules.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.590950  DOI: Not available
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