The Interaction and Expression Analysis of OsSH3P2 in Rice (Oryza sativa)
WU Zhi-Yuan1,2,*, XIE Yun-Jie1,2,*, WANG Yu-Peng1,2, XUE Chen-Hao1,2, WU Xing-Qian1,2, LAI Yu-Ling1,2, WEI Jing-Wen1,2, CAI Qiu-Hua1,2, XIE Hua-An1,2, ZHANG Jian-Fu1,2,**
1 Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350019, China; 2 Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China, Ministry of Agriculture and Rural Affairs/Fuzhou Branch, National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/Fujian Key Laboratory of Rice Molecular Breeding/Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding Between Fujian and Ministry of Sciences & Technology/Base of South China, National Key Laboratory of Hybrid Rice/National Rice Engineering Laboratory of China, Fuzhou 350003, China
Abstract：SH3 domain-containing proteins (SH3Ps) in plant are members of a broad protein category and have been proved to be involved in several processes in cells. In Arabidopsis thaliana, the interaction network of SH3P2 is extensive and plays an important role in endocytosis, vesicle trafficking, binding ubiquitin, cell division and autophagosome formation. Up to now, little has been reported about SH3P2 in rice (Oryza sativa). In this study, the rice OsSH3P2 gene was introduced into protoplasts for transient expression. It was observed that OsSH3P2 was mainly localized in clathrin-coated vesicle, and partially localized in the trans-Golgi network/early endosome. Meanwhile, the subcellular localization results showed that OsSH3P2 could co-localize with protein ELC-like (ELC), a member of vesicle transport complex. Further, various means such as yeast two-hybrid (Y2H), co-immunoprecipitation (Co-IP) and bimolecular fluorescence complementation (BiFC) were used to confirm the interaction between OsSH3P2 and ELC, indicating they were highly correlated. Subsequently, by extracting the RNA from various tissues of rice and conducting qRT-PCR detection, the results showed that the expression level of OsSH3P2 was relatively stable at all stages of rice growth, and was the highest in stem. The PlantCARE online analysis revealed that there were multiple light-responsive regulatory elements in the promoter region of OsSH3P2. A dual-luciferase reporter assay was designed to confirm that the expression of OsSH3P2 was affected by light and was induced under dark conditions. This study revealed the cellular localization, protein interaction and expression pattern of OsSH3P2 in rice, laying a preliminary theoretical foundation for further exploration of the biological functions of OsSH3P2.
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