The cowpea RING ubiquitin ligase VuDRIP interacts with transcription factor VuDREB2A for regulating abiotic stress responses

•We cloned the DREB2A-interacting protein (VuDRIP) homolog of drought-tolerant plant cowpea by degenerate PCR and RACE approaches.•We studied the expression of the VuDRIP gene under different abiotic stress and phytohormones by quantitative PCR.•We studied interaction of VuDRIP with VuDREB2A by the yeast two-hybrid system.

Cowpea (Vigna unguiculata L. Walp) is an important grain legume cultivated in drought-prone parts of the world, having higher tolerance to heat and drought than many other crops. The transcription factor, Dehydration-Responsive Element-Binding protein 2A (DREB2A), controls expression of many genes involved in osmotic and heat stress responses of plants. In Arabidopsis, DREB2A-interacting proteins (DRIPs), which function as E3 ubiquitin ligases (EC 6.3.2.19), regulate the stability of DREB2A by targeting it for proteasome-mediated degradation. In this study, we cloned the cowpea ortholog of DRIP (VuDRIP) using PCR based methods. The 1614 bp long VuDRIP mRNA encoded a protein of 433 amino acids having a C3HC4-type Really Interesting New Gene (RING) domain in the N-terminus and a C-terminal conserved region, similar to Arabidopsis DRIP1 and DRIP2. We found VuDRIP up-regulation in response to various abiotic stresses and phytohormones. Using yeast (Saccharomyces cerevisae) two-hybrid analysis, VuDRIP was identified as a VuDREB2A-interacting protein. The results indicate negative regulation of VuDREB2A by ubiquitin ligases in cowpea similar to Arabidopsis along with their other unknown roles in stress and hormone signaling pathways.