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Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
DNA binding-induced conformational change of the yeast transcriptional activator PRTF
1Institut für Molekularbiologie und Biophysik, Eth-Hönggerberg, Zurich, Switzerland.
Abstract:
Our studies using proteases to probe protein structure establish that binding to the upstream activating sequences (UASs) of two different yeast a-specific genes induces a conformational change in the pheromone/receptor transcription factor (PRTF), which is not observed upon binding to the UASs of either of two alpha-specific genes. We propose that this selective structural alteration exposes an activation region of PRTF when it binds a-specific genes, switching these genes on. The transcriptional activator MAT alpha 1 may activate alpha-specific genes by binding to the PRTF-alpha-specific UAS complex and unmasking the otherwise hidden activation surface of PRTF. We also show that the N-terminal third of PRTF is sufficient for specific DNA binding, while the middle third of the protein interacts with MAT alpha 1.
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