The TIR-NBS 2 (TN2) gene from Arabidopsis thaliana (Arabidopsis), which encodes a TIR (the Toll and Interleukin-1 Receptor)-type of nucleotide binding site (NBS) receptor protein (TIR-NBS) that can cause cell death in the model plant Nicotiana benthamiana (N. benthamiana). Nevertheless, the mechanism of TN2 signal initiation is still unclear. This research performed yeast two-hybrid and bimolecular fluorescence complementation (BIFC) assays to investigate interactions between proteins of TN2, and analyzed the influences of these interactors on TN2 function using N. benthamiana. EXO70B1, SOC3 and CPK5-VK were identified as interacting proteins of TN2 based on yeast two-hybrid and BIFC methods. Functional annotations of these interacting proteins indicate their involvement in multiple pathways, including exocytosis, positive regulation of abscisic acid-activated signaling pathway, regulation of stomatal closure, response to water deprivation, defense response, signal transduction and intracellular signal transduction. The transient assay results proclaimed that EXO70B1 can suppress cell death triggered by TN2 and TN2-TIR. These outcomes suggest that TN2 receptor may be participated in various pathways, and the protein level and activity are strictly controlled at multiple aspects, providing novel clues for elucidating the molecular mechanism of TN2 immune receptor in Arabidopsis resistance.
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