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Protein protein and dna protein transactions during gene activation activator sigma n contacts and the dna melting activity of the sigma n activator complex

Objective



Determination of the critical protein-protein interactions in transcription systems and in particular between sigma-N and its activator, and the contribution of the sigma-N holoenzyme-activator complex to the DNA strand separation (melting) process are essential for the understanding of the mechanism of transcriptional activation of the RNA polymerase holoenzyme. The sigma-N protein is well suited to analysis as biochemically active domains have been isolated. The overall aims of the proposed work are to: (a) Locate the putative protein-protein interactions between activator and the sigma-N protein, that appear to be critical for activation of RNA polymerase holoenzyme.
(b) Identify promoter DNA-contact regions of the sigma-N
holoenzyme-activator complex during catalysis of open complex formation. (c) Mutate candidate DNA and activator contact regions of sigma-N identified as described above. Screen mutants defective in either a DNA contact during DNA melting and/or activation interaction.
(d) Analyse the DNA-melting process, dependent of the formation of the sigma-N holoenzyme-activator complex.

Funding Scheme

RGI - Research grants (individual fellowships)

Coordinator

IMPERIAL COLLEGE OF SCIENCE, TECHNOLOGY AND MEDICINE
Address
Campus At Silwood Park, Road A329 (Virginia Water
SW7 2AZ London
United Kingdom

Participants (1)

Not available
Spain