Screening DHHCs of S‐acylated proteins using an OsDHHC cDNA library and bimolecular fluorescence complementation in rice. (11th May 2022)

• Record Type: Journal Article
• Authors: Tian, Ye; Zeng, Hui; Wu, Jicai; Huang, Jian; Gao, Qiang; Tang, Dongying; Cai, Lipeng; Liao, Zhaoyi; Wang, Yan; Liu, Xuanming; Lin, Jianzhong
• Is Part Of: Plant journal. Volume 110:Number 6(2022)
• Page Start: 1763
• Page End: 1780
• Publication Date: 2022-05-11
• Subjects: bimolecular fluorescence complementation -- OsDHHC30 -- S‐acylation -- OsCBL2/3 -- rice (Oryza sativa L.) -- OsDHHC cDNA library; Plant molecular biology -- Periodicals; Plant cells and tissues -- Periodicals; Botany -- Periodicals; 580
• DOI: 10.1111/tpj.15769
• Languages: English
• ISSNs: 0960-7412

Abstract

SUMMARY: S ‐acylation is an important lipid modification that primarily involves DHHC proteins (DHHCs) and associated S ‐acylated proteins. No DHHC– S ‐acylated protein pair has been reported so far in rice ( Oryza sativa L.) and the molecular mechanisms underlying S ‐acylation in plants are largely unknown. We constructed an OsDHHC cDNA library for screening corresponding pairs of DHHCs and S ‐acylated proteins using bimolecular fluorescence complementation assays. Five DHHC– S ‐acylated protein pairs (OsDHHC30–OsCBL2, OsDHHC30–OsCBL3, OsDHHC18–OsNOA1, OsDHHC13–OsNAC9, and OsDHHC14–GSD1) were identified in rice. Among the pairs, OsCBL2 and OsCBL3 were S ‐acylated by OsDHHC30 in yeast and rice. The localization of OsCBL2 and OsCBL3 in the endomembrane depended on S ‐acylation mediated by OsDHHC30. Meanwhile, all four OsDHHCs screened complemented the thermosensitive phenotype of an akr1 yeast mutant, and their DHHC motifs were required for S ‐acyltransferase activity. Overexpression of OsDHHC30 in rice plants improved their salt and oxidative tolerance. Together, these results contribute to our understanding of the molecular mechanism underlying S ‐acylation in plants. Significance Statement: S ‐acylation is an important lipid modification that involves DHHCs and S ‐acylated proteins, but DHHC– S ‐acylated protein pairs have not been reported in rice ( Oryza sativa L.). In this study, an OsDHHC cDNA library was constructed to screen the DHHCs of S ‐acylated proteins using a bimolecular fluorescence complementation assay, which led to the identification of five DHHC– S ‐acylated protein pairs in rice.