Homo-dimerization and ligand binding by the leucine-rich repeat domain at RHG1/RFS2 underlying resistance to two soybean pathogens.
Homo-dimerization and ligand binding by the leucine-rich repeat domain at RHG1/RFS2 underlying resistance to two soybean pathogens.
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DOI：
10.1186/1471-2229-13-43
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发表时间：
2013-03-15
影响因子：
5.3
通讯作者：
Lightfoot DA
中科院分区：
文献类型：
--
作者：
Afzal AJ;Srour A;Goil A;Vasudaven S;Liu T;Samudrala R;Dogra N;Kohli P;Malakar A;Lightfoot DA
由GmRLK 18 -1（18号染色体上的Glyma_18_02680）编码的蛋白质是在大豆（Glycine max L.）梅尔）Rhg 1/Rfs 2基因座。该基因座是大豆胞囊线虫（Heterodera glycines（I.）以及猝死综合征（SDS）的病原体大豆球孢镰刀菌（Fusarium virguliforme（Aoki））。以前，富含亮氨酸重复序列（LRR）结构域在大肠杆菌中表达。本文的目的是评估LRR的能力;同源二聚化;结合较大的蛋白质;和结合小肽。Western分析表明，从根中提取蛋白质后可以形成同源二聚体。观察到来自残基131-485的纯化的LRR结构域在体外形成单体和同源二聚体的混合物。体外交联实验显示H274 N区域接近（<11.1 A）第二同源二聚体亚基上高度保守的半胱氨酸残基C196。发现在植物和线虫分泌物中发现的肽的结合常数为20-142 nM。当用50 pM的肽处理根时，观察到对植物表型的影响，包括萎蔫、茎弯曲和对SCN感染的抗性。远Western分析，随后MS显示甲硫氨酸合酶和亲环蛋白强烈结合LRR结构域。来自GmRLK 08 -1（Glyma_08_g11350）的第二个LRR未显示这些强相互作用。GmRLK 18 -1蛋白的LRR结构域形成单体和同源二聚体。LRR结构域与4种不同的CLE肽、亲环蛋白和甲硫氨酸合酶强烈结合。CLE肽GmTGIF、GmCLE 34、GmCLE 3和HgCLE先前被报道参与根生长抑制，但这里GmTGIF和HgCLE显示改变茎形态和对SCN的抗性。从同源性和从头建模的几个模型之一，部分通过交联验证。预测RLK同种异型A87 V、Q115 K和H274 N中存在的3个氨基酸置换的作用改变结构域稳定性和功能。因此，GmRLK 18 -1的LRR结构域可能是大豆根系发育和抗病性的基础，并提供了开发新的变体和配体的途径，这些变体和配体可能促进减少SCN的损失。
The protein encoded by GmRLK18-1 (Glyma_18_02680 on chromosome 18) was a receptor like kinase (RLK) encoded within the soybean (Glycine max L. Merr.) Rhg1/Rfs2 locus. The locus underlies resistance to the soybean cyst nematode (SCN) Heterodera glycines (I.) and causal agent of sudden death syndrome (SDS) Fusarium virguliforme (Aoki). Previously the leucine rich repeat (LRR) domain was expressed in Escherichia coli. The aims here were to evaluate the LRRs ability to; homo-dimerize; bind larger proteins; and bind to small peptides. Western analysis suggested homo-dimers could form after protein extraction from roots. The purified LRR domain, from residue 131–485, was seen to form a mixture of monomers and homo-dimers in vitro. Cross-linking experiments in vitro showed the H274N region was close (<11.1 A) to the highly conserved cysteine residue C196 on the second homo-dimer subunit. Binding constants of 20–142 nM for peptides found in plant and nematode secretions were found. Effects on plant phenotypes including wilting, stem bending and resistance to infection by SCN were observed when roots were treated with 50 pM of the peptides. Far-Western analyses followed by MS showed methionine synthase and cyclophilin bound strongly to the LRR domain. A second LRR from GmRLK08-1 (Glyma_08_g11350) did not show these strong interactions. The LRR domain of the GmRLK18-1 protein formed both a monomer and a homo-dimer. The LRR domain bound avidly to 4 different CLE peptides, a cyclophilin and a methionine synthase. The CLE peptides GmTGIF, GmCLE34, GmCLE3 and HgCLE were previously reported to be involved in root growth inhibition but here GmTGIF and HgCLE were shown to alter stem morphology and resistance to SCN. One of several models from homology and ab-initio modeling was partially validated by cross-linking. The effect of the 3 amino acid replacements present among RLK allotypes, A87V, Q115K and H274N were predicted to alter domain stability and function. Therefore, the LRR domain of GmRLK18-1 might underlie both root development and disease resistance in soybean and provide an avenue to develop new variants and ligands that might promote reduced losses to SCN.
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影响因子：
64.8
作者：
Demarest, SJ;Martinez-Yamout, M;Wright, PE
通讯作者：
Wright, PE
影响因子：
6.8
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