RNA-UNY 2010 program

ERI Mediated Decay (EMD): Novel exonucleases and pathways in mRNA decay

Nithya Krishnan, Anand Parekh, Patrick Itotia, MingJing Wu, and Roopa Thapar

Hauptman-Woodward Medical Research Institute and Department of Structural Biology, SUNY, Buffalo

The ERI family of nucleases are involved in a spectrum of protein-protein and protein-nucleic acid interactions and have been implicated in an array of cellular processes: histone mRNA metabolism, RNAi, 5.8S rRNA processing, and miRNA mediated translational repression and/ or decay. ERI-2 or Snipper (in Drosophila) are uncharacterized members of the DEDDh subfamily of exonucleases. We recently published that Snipper1 is a highly active 35 exonuclease in vitro and can act on both linear and double-stranded RNA substrates. A biochemical property shared by the ERI family of nucleases is that they prefer double- stranded RNA substrates with short 3 overhangs of 2-5 nucleotides, reminiscent of siRNAs/ miRNAs or stem-loop structures. Ongoing studies in our laboratory indicate that hERI-2 may be a RNA-Induced Silencing Complex (RISC) associated nuclease that regulates polyA binding protein (PABP)-dependent mRNA turnover by promoting micro-RNA mediated mRNA decay. hERI-2 interacts directly with PABPC1 in vitro and in vivo. hERI-2 also co-localizes with Ago2 but is not present in P-bodies. Microarray analysis of hERI-2 siRNA treated cells shows that it is involved in regulating the stability of a number of cell cycle regulators, chemokines and cytokines, particularly those with AU-rich elements in their 3 untranslated regions. Three mRNAs implicated in disease states that are targeted by hERI-2 are the cyclin inhibitor and tumor suppressor CDKN1A, the chemokine CCL5, and the cytokine IL-8. ERI-2 also regulates the cell cycle. Ongoing studies on both ERI-1 and ERI-2 to understand the functional and structural roles of these nucleases in mRNA decay of specific mRNA targets will be presented.

1.Kupsco, J.M., Wu, M.J., Marzluff, W.F., Thapar, R., and Duronio, R.J. Genetic and biochemical characterization of Drosophila Snipper: A promiscuous member of the metazoan 3hExo/ERI-1 family of 3 to 5 exonucleases. RNA (2006), 12(12): 2103.

RNA-UNY 2010 program