The Serum Response Element (SRE) is an essential regulatory element found in the promoters of many genes, including c-fos, whose transcription is rapidly and transiently induced following growth factor or mitogen stimulation. The transcription factor known as Serum Response Factor (SRF) and its accessory protein, Ternary Complex Factor (TCF), bind SRE cooperatively to form a ternary complex. Activation of SRE by TCF-linked signal pathways is relatively well understood. In addition, SRF can activate transcription independently of TCF via a novel signal transduction pathway (SRF linked signal pathway). The objective of the proposal is to characterise this pathway. This will be done using four different approaches:
- Characterisation of stimuli and inducers that activate the SRF-linked signal pathway (Investigating M1 and M2 muscarinic receptors as activators of this pathway; finding agents inducing specifically this pathway). - Several candidate regulatory factors (members of different kinase families) will be tested as activators of the SRF-linked pathway. - Identification of potential kinases involved the SRF-linked signal pathway will be tried by a series of biochemical methods.
- Cloning cDNAs that activate the SRF-linked signal pathway will be attempted using a functional cDNA library screening method. The post-doctorate will be trained in many advanced molecular biology techniques (transfection, cDNA library screening, gene expression analysis). Training will also contain all the biochemical techniques necessary for the analysis of signalling molecules, microinjection of DNAs and proteins, FACS (Fluorescence Activated Cell Sorting).