SWI3C-CRCS · Characterization of signalling functions and genomic targets of SWI3C-associated chromatin remodelling complexes

Chromatin-remodelling complexes (CRCs) play pivotal roles in the regulation of basic cellular processes, including transcription, DNA replication, repair and cell cycle in eukaryotes. Although several orthologues of CRC components are conserved between higher plants and other eukaryotes, our current knowledge on plant CRCs is rather limited. To unravel the regulatory functions of SWI/SNF-type CRCs, the proposed study focuses on functional analysis of SWI3 core subunits that are linked by ATPases and other interacting factors to specific signalling pathways. Recently, we demonstrated that SWI/SNF CRCs in Arabidopsis carry different combinations of SWI3 subunits that play distinct and essential roles in the control of cell differentiation, organ development, flowering time and gene silencing. The SWI3 subunits show unique interactions with several signalling factors, such as the flowering time regulator FCA. Through their SNF5/BSH CRC subunit, the CRC complexes interact with conserved components of stress signalling pathways, including the AMPK-activated protein kinases. The proposed training project is based on a focused multidisciplinary approach, which aims at the characterization of regulatory roles of SWI/SNF CRC complexes that carry ATSWI3C subunit. Complementary tasks of the experimental work plan include: i) the isolation and mass spectrometry analysis of CRC complexes carrying affinity tag labeled SWI3C subunit, ii) identification of genomic targets of SWI3C containing CRCs by chromatin cross-linking, iii) analysis of function of SWI3C-associated CRC components using reverse genetics, and iv) study of in vivo nuclear interaction of CRC subunits using fluorescence energy transfer-based confocal microscopy. The project is aided by expert mentoring activity of the host and is embedded in a carrier development plan envisioning long-term collaborative interaction with the researcher upon his return to the home institute.