RRM
2010 Rustbelt RNA Meeting
RRM
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Poster abstracts
Abstract:
Polyadenylation is one of the most important post-transcriptional processes that result in the formation of mature mRNA. Identification of specific regions on RNA, binding to the RNA and its interaction with protein complexes that associate with other protein complexes play key role in 3’-end processing. In plants, this process is guided by Cis-regulatory elements like far-upstream element (FUE), near-upstream elements (NUE) and cleavage/ polyadenylation site. The multi-subunit protein complexes like Cleavage and polyadenylation specificity factor (CPSF), Cleavage stimulation factor (CstF), Cleavage factors and Poly(A) Polymerase are important factors for polyadenylation in eukaryotes. In Mammals, the Cleavage stimulation factor (CstF) is made up of three subunits CstF 77, CstF64 and CstF 50.
Here we focus on Arabidopsis (FY) orthologue of the essential yeast subunit Pfs2p. FY (At5g13480) is a known 3’-end processing factor and plays a key role in the auto regulation of FCA and interacts with FCA for chromatin silencing that targets FLC (flowering locus C), a repressor of flowering. We established a tethering assay to study the role of FY as a general 3’-end processing factor using transients assays by Agro-infiltrations. Initial results using this assay suggest that tethering of FY to an RNA does not promote polyadenylation of the RNA. Studies are under way to confirm these results, using 3’-RACE and tethering assays involving other polyadenylation factor subunits.
FY also interacts with other polyadenylation factors like CstF77, CPSF 100, CPSF160, CPSF 30 and Fip1 in carrying out the polyadenylation reaction. Our hypothesis is that if FY is unable to bring 3’-end processing complex to the RNA via tethering, it might be one of these other proteins that accomplish this task.
References:
1. Li, Q. and A. G. Hunt (1997). The Polyadenylation of RNA in Plants. 115: 321-325.
2.
Y Takagaki, J L Manley, C C MacDonald, J Wilusz, and T Shenk (1990) A multisubunit factor, CstF, is required for polyadenylation of mammalian pre-mRNAs.
3. Richard M. Amasino (2003). Flowering time: a pathway that begins at the 3 end. vol 13 Issue 17.
4. David Manzano, Sebastian Marquardt, Alexandra M. E. Jones, Isabel Bäurle, Fuquan Liu, and Caroline Dean-Altered interactions within FY/AtCPSF complexes required for Arabidopsis FCA-mediated chromatin silencing.PNAS 2009 106 (21) 8772-8777;
5. Arthur G Hunt*1, Ruqiang Xu2,3, Balasubrahmanyam Addepalli1,Suryadevara Rao1, Kevin P Forbes1,4, Lisa R Meeks1,5, Denghui Xing2,Min Mo2,6, Hongwei Zhao2, Amrita Bandyopadhyay1,Lavanya Dampanaboina1, Amanda Marion1, Carol Von Lanken1 and Qingshun Quinn Li*2.BMC Genomics 2008, 9:220 doi:10.1186/1471-2164-9-220
Keywords: Tethering, RACE (Rapid Amplification of cDNA Ends), Polyadenylation
