Arabidopsis thaliana PRP40s are RNA polymerase II C-terminal domain-associating proteins |
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Authors: | Chang Ho Kang Yue Feng In Sil Jeong Jeong Dong Bahk Sang Yeol Lee Hisashi Koiwa |
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Institution: | a Vegetable and Fruit Improvement Center, Department of Horticultural Sciences, and Molecular and Environmental Plant Science Program, Texas A&M University, College Station, TX 77843-2133, USA b Division of Applied Life Science (BK21 Program) and Environmental Biotechnology National Core Research Center, Graduate School of Gyeongsang National University, Jinju 660-701, South Korea |
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Abstract: | The carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II functions as a scaffold for RNA processing machineries that recognize differentially phosphorylated conserved (YSPTSPS)n repeats. Evidence indicates that proteins that regulate the phosphorylation status of the CTD are determinants of growth, development, and stress responses of plants; however, little is known about the mechanisms that translate the CTD phosphoarray into physiological outputs. We report the bioinformatic identification of a family of three phospho-CTD-associated proteins (PCAPs) in Arabidopsis and the characterization of the AtPRP40 (Arabidopsis thaliana PRE-mRNA-PROCESSING PROTEIN 40) family as PCAPs. AtPRP40s-CTD/CTD-PO4 interactions were confirmed using the yeast two-hybrid assay and far-Western blotting. WW domains at the N-terminus of AtPRP40b mediate the AtPRP40b-CTD/CTD-PO4 interaction. Although AtPRP40s interact with both phosphorylated and unphosphorylated CTD in vitro, there is a strong preference for the phosphorylated form in Arabidopsis cell extract. AtPRP40s are ubiquitously expressed and localize to the nucleus. These results establish that AtPRP40s are specific PCAPs, which is consistent with the predicted function of the AtPRP40 family in pre-mRNA splicing. |
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Keywords: | CTD carboxyl-terminal domain RNA polymerase II RNA processing machineries CTD phosphoarray PCAPs phospho-CTD-associated proteins AtPRP40 arabidopsis thaliana PRE-mRNA-PROCESSING PROTEIN 40 Yeast two-hybrid assay WW domains pre-mRNA splicing In silico search |
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