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J Exp Bot


Title:The type 3 effector NopL of Sinorhizobium sp. strain NGR234 is a mitogen-activated protein kinase substrate
Author(s):Ge YY; Xiang QW; Wagner C; Zhang D; Xie ZP; Staehelin C;
Address:"State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, Guangzhou, China. State Key Laboratory of Biocontrol and Guangdong Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-sen University, Guangzhou, China Shenzhen Research and Development Center of State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Baoan, Shenzhen, China xiezping@mail.sysu.edu.cn cst@mail.sysu.edu.cn"
Journal Title:J Exp Bot
Year:2016
Volume:20160229
Issue:8
Page Number:2483 - 2494
DOI: 10.1093/jxb/erw065
ISSN/ISBN:1460-2431 (Electronic) 0022-0957 (Linking)
Abstract:"Pathogenic bacteria utilize type 3 secretion systems to inject type 3 effectors (T3Es) into host cells, thereby subverting host defense reactions. Similarly, T3Es of symbiotic nitrogen-fixing rhizobia can affect nodule formation on roots of legumes. Previous work showed that NopL (nodulation outer protein L) of Sinorhizobium(Ensifer) sp. strain NGR234 is multiply phosphorylated in eukaryotic cells and that this T3E suppresses responses mediated by mitogen-activated protein (MAP) kinase signaling in yeast (mating pheromone signaling) and plant cells (expression of pathogenesis-related defense proteins). Here, we show that NopL is a MAP kinase substrate. Microscopic observations of fluorescent fusion proteins and bimolecular fluorescence complementation analysis in onion cells indicated that NopL is targeted to the nucleus and forms a complex with SIPK (salicylic acid-induced protein kinase), a MAP kinase of tobacco. In vitro experiments demonstrated that NopL is phosphorylatyed by SIPK. At least nine distinct spots were observed after two-dimensional gel electrophoresis, indicating that NopL can be hyperphosphorylated by MAP kinases. Senescence symptoms in nodules of beans (Phaseolus vulgaris cv. Tendergreen) were analyzed to determine the symbiotic effector activity of different NopL variants with serine to alanine substitutions at identified and predicted phosphorylation sites (serine-proline motif). NopL variants with six or eight serine to alanine substitutions were partially active, whereas NopL forms with 10 or 12 substituted serine residues were inactive. In conclusion, our findings provide evidence that NopL interacts with MAP kinases and reveals the importance of serine-proline motifs for effector activity during symbiosis"
Keywords:Bacterial Proteins/*metabolism Cell Nucleus/metabolism MAP Kinase Signaling System Mitogen-Activated Protein Kinases/*metabolism Mutation/genetics Phaseolus/physiology Phosphorylation Plant Root Nodulation Protein Binding Saccharomyces cerevisiae/metaboli;
Notes:"MedlineGe, Ying-Ying Xiang, Qi-Wang Wagner, Christian Zhang, Di Xie, Zhi-Ping Staehelin, Christian eng Research Support, Non-U.S. Gov't England 2016/03/05 J Exp Bot. 2016 Apr; 67(8):2483-94. doi: 10.1093/jxb/erw065. Epub 2016 Feb 29"

 
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