Molecular analysis of the mechanisms linking co-transcriptional RNA processing with chromatin silencing

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

The project will exploit our recent work demonstrating that FCA, an Arabidopsis protein containing RNA-binding domains (RRM) mediates repression of a floral repressor gene FLC through a transcriptional silencing mechanism. Our aim will be to dissect how RNA metabolism drives chromatin changes, an important question in many areas of biology. Chromatin immunoprecipitation experiments will be used to explore the relationship between RNA processing and recruitment of FCA to FLC. We will characterize in an unbiased way all the FLC sense and antisense transcripts in different genotypes and define the RNA species to which FCA binds in vivo. In order to establish the generality of our findings we will also explore the genome-wide RNA features recognized by FCA and work collaboratively with Dr H. Chang (Stanford University) who is exploring how non-coding RNAs drive chromatin changes at mammalian HOX loci. Chromatin immunoprecipitation will be used to explore the requirements for FLD association to FLC chromatin and determine if the association of the chromatin regulator, FLD, to its target FLC is dependent on the RNA metabolic activities of FCA and FY. Working collaboratively with Prof. Y. Shi (Harvard Medical School - who first demonstrated histone demethylases exist) we will test the generality of RRM protein recruitment of histone demethylases to their targets by analysing the role of the most homologous human RRM proteins to FCA, on LSD1 targets in human cells. Lastly, in order to mechanistically understand the connection between chromatin modification activity and factors involved in RNA processing we will use tandem affinity purification to identify FLD associated proteins and RNA in vivo.

unavailable