Arabidopsis Polyadenylation Factor Subunits - Mutants and Protein Interaction Networks

The polyadenylation of messenger RNAs (mRNAs) is an important step in the process of gene expression in plants. Polyadenylation adds an essential element to the mRNA and helps to coordinate the synthesis of a messenger RNA with its ultimate functioning in protein synthesis. Consequently, the protein subunits that make up the polyadenylation machinery are expected to play important roles in plant growth. The goals of this project are to determine the collective function of the set of Arabidopsis genes that encode subunits of the polyadenylation machinery (this set, including Arabidopsis genome identifiers, can be found at http://www.uky.edu/~aghunt00/polyA2010.html and http://www.users.muohio.edu/liq/arabidopsis2010.html). This will include a documentation of the phenotypes of plants that have had one or more subunit removed (by mutation or gene silencing) and a determination of the complete set of protein-protein interactions involving the various subunits. The results will be disseminated via peer-reviewed publication as well as through web pages managed by the principal investigators (http://www.uky.edu/~aghunt00/polyA2010.html and http://www.users.muohio.edu/liq/arabidopsis2010.html). In addition, mutant Arabidopsis stocks will be deposited with the appropriate stock center(s). The research will contribute to a goal of the 2010 Project ("to determine the function of all genes of a reference species within their cellular, organismal and evolutionary contexts by the year 2010") by cataloguing mutant phenotypes, by defining the network of interactions amongst Arabidopsis polyadenylation factor subunits, and by identifying proteins of unknown function that interact with the polyadenylation machinery. The research will greatly increase the current understanding of mRNA 3' end formation in higher plants. As such, it will enhance the ability to tailor foreign genes for expression in transgenic plants, and thereby develop improved crops (as well as novel alternative economic opportunities) for farmers. It will also lend significant insight into the evolution of mRNA polyadenylation. The research products (cloned cDNA, antibodies, plant lines etc.) will be made available to the research community at large. The research will contribute to the training of two postdoctoral scholars, four graduate students, and as many as twelve undergraduates. The latter will be drawn from highly-regarded and diverse programs at the University of Kentucky and Miami University.