Towards the identification of Tudor SN interactome : cloning of putative interactors

Abstract: Tudor staphylococcal nuclease (TSN) is a protein found associated with stress granules (SG) and processing bodies (PB) where it acts as a part of the plant stress response. In plants not much is known for TSN. This project serves as a starting point in our attempt to elucidate the physically interacting proteins of TSN. Previously, our group identified proteins that can be physical interactors of TSN. Herein, we focused on TCTP, MAPK, Armadillo, FY, DHH1 and Kin11. By using overlay PCR, I fused the cDNAs of these genes with their corresponding promoters, and by using the Gateway technology I finally introduced these fusions to a binary vector which contains the protein fluorescent marker GFP. These binary vectors were transformed into Agrobacterium tumefaciens. In parallel, the promoters of these genes were introduced into a binary vector upstream of a uidA gene (encoding β-glucoronidase; GUS). In the future the aforementioned transformed Agrobacterium cells will be used to transform Arabidopsis plants to confirm the interactions of the above mentioned proteins with TSN and its association with SG and PB.