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Definition of the protein components of the FCA/FY complex in Arabidopsis thaliana (FCA/FY_complex)

Reference	BBS/E/J/0000A219
Principal Investigator / Supervisor	Professor Dame Caroline Dean
Co-Investigators / Co-Supervisors
Institution	John Innes Centre
Department	John Innes Centre Department
Funding type	Research
Value (£)	57,151
Status	Completed
Type	Institute Project
Start date	01/01/2005
End date	31/12/2006
Duration	24 months

Abstract

The autonomous promotion pathway plays a major role in controlling the floral transition in Arabidopsis. Its main function is to down-regulate RNA levels of the floral repressor, FLC. The Dean lab has cloned two components of this pathway, FCA and FY, which encode an RNA-binding protein and a homologue of a yeast protein required for mRNA 3¿-end-processing and polyadenylation respectively. The Dean lab has recently shown that FCA/FY proteins interact physically and regulate the alternative processing of the FCA transcript. Since FCA has no homology to any known component of the core polyadenylation machinery, it might be a regulator of 3¿-end formation, whereas FY (by sequence similarity) seems to be a conserved component of the 3¿-end processing machinery. This finding is of great interest because although alternative polyadenylation is increasingly well documented and can have profound effects on gene expression, little is known about trans-acting factors that regulate it and no tissue-specific or transcript-specific factors have been described. The characterization of Arabidopsis FCA/FY complex will enable this question to be addressed, and will help to elucidate general questions about the composition of complexes regulating RNA 3¿-end formation in eukaryotes. The FY yeast homologue (Pfs2p) interacts with Ysh1p, Fip1p, and RNA14p. We shall first clone Arabidopsis orthologues of Ysh1p, Fip1p, and RNA14p, and we will test if the FY protein interacts with them by GST pull-down assay. Secondly, we will also identify other components of the FCA/FY complex by yeast two-hybrid experiments using FY as bait. Finally, we will express FCA and FY-tagged proteins with TAP and HA epitopes in cultured Arabidopsis cells and in Arabidopsis transgenic plants for the purification of the FCA/FY complex by affinity chromatography. We will identify these components by multidimensional protein identification technology.

Summary

unavailable

Committee	Closed Committee - Genes & Developmental Biology (GDB)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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