| Summary: | The SWI/SNF complex is a large, multi subunit chromatin-remodelling transcriptional regulator complex with an ATPase subunit, Brahma. Recently published data from our laboratory has revealed that Brahma regulates the activity of the EGFR signalling pathway in Drosophila. The study presented here was conducted to further elucidate which components of the EGFR/Ras/Raf/Erk pathway are regulated by Brahma, and also the mechanism behind this regulation. Analysis of components in this pathway at both the transcriptional and protein levels following overexpression of wild-type Brahma in Drosophila S2 cells, demonstrated that Brahma may affect EGFR mRNA expression and Ras protein level. In addition, previous studies suggest that Brahma activity is regulated through phosphorylation by kinases, including Erk and cyclin E/CDK 2. Analysis of Brahma immunoprecipitated from S2 cells by mass spectrometry revealed contamination by Myosin. Ion exchange chromatography and ammonium sulphate precipitation were used to separate Brahma from Myosin for phosphorylation site analysis by mass spectrometry.
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