RRM
2010 Rustbelt RNA Meeting
RRM
|
|
|
|
|
Poster abstracts
Abstract:
CPSF30 is a component in 3’ RNA processing that has been well characterized in the model species Arabidopsis thaliana (1). In this study the gene for CPSF30 has been cloned out of Medicago truncatula and Medicago sativa. A large region within the gene exhibits significant conservation between A. thaliana and both Medicago species. This area of the gene was cloned out and 5’ and 3’ RACE were used to get each full length gene. The Medicago genes were cloned into a protein expression vector. The vector was used to translate each gene into its corresponding protein and each protein was purified. CPSF30 from A. thaliana has been shown to possess three CCCH type zinc fingers, with the first implicated in RNA binding and the third in RNA cleavage (1). Amino acid alignments showed the zinc finger regions were conserved and it was hypothesized that CPSF30 from both Medicago species would bind RNA and cleave RNA. Electromobility shift assays were used to determine that RNA binding did occur. RNA nuclease activity in Medicago is still being tested. CPSF30 knockout studies in A. thaliana have yielded some consistent phenotypes that are still being characterized and explored. One such characteristic is an elevated resistance to oxidative stress (2). CPSF30 knockouts have been produced in Medicago sativa. Preliminary data suggests that the absence of CPSF30 expression results in a distinct root phenotype. The CPSF30 knockout plants were crossed with a cultivar, Spreader4, which is an agriculturally relevant variety. It has been determined that the knockout vector is present in the hybrid progeny and phenotypes are still being evaluated.
References:
(1) Delaney KJ, Xu R, Zhang J, Li QQ, Yun KY, et al. (2006) Calmodulin interacts with and regulates the RNA-binding activity of an Arabidopsis polyadenylation factor subunit. Plant Physiol 140: 1507–1521.
(2) Zhang J, Addepalli B, Yun K-Y, Hunt AG, Xu R, et al. (2008) A Polyadenylation Factor Subunit Implicated in Regulating Oxidative Signaling in Arabidopsis thaliana. PLoS ONE 3(6): e2410. doi:10.1371/journal.pone.0002410
Keywords: CPSF30, Medicago
