<br>由分子内相互作用调节的 Pm21 CC 结构域活性与细胞死亡和抗病性有关。,Molecular Plant Pathology

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由分子内相互作用调节的 Pm21 CC 结构域活性与细胞死亡和抗病性有关。
Molecular Plant Pathology ( IF 4.8 ) Pub Date : 2020-05-18 , DOI: 10.1111/mpp.12943
Anli Gao ₁ , Minjie Hu ₁ , Yifei Gong ₁ , Ruixiang Dong ₁ , Yuan Jiang ₁ , Shanying Zhu ₂ , Jian Ji ₂ , Dale Zhang ₁ , Suoping Li ₁ , Huagang He ₂

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核苷酸结合 (NB) 富含亮氨酸重复序列 (LRR) 受体 (NLR) 可抵抗多种植物病原体。我们之前克隆了小麦白粉病抗性基因Pm21 ，该基因编码卷曲螺旋 (CC) NLR，赋予对Blumeria graminis f 的广谱抗性。 sp 。小麦。在这里，我们报告了本塞姆氏烟草中 Pm21 CC 结构域的全面生化和功能分析。瞬时过表达测定表明，只有扩展的 CC（eCC，氨基酸残基 1-159）结构域具有细胞死亡诱导活性，而包含 CC 的截短片段，包括 CC-NB 和 CC-NB-LRR，不会诱导细胞死亡。 ‐死亡反应。免疫共沉淀 (Co-IP) 测定表明，eCC 结构域与植物中的 NB 和 LRR 结构域自关联并相互作用。这些结果表明，在没有病原体的情况下，eCC 结构域的活性受到不同结构域的分子内相互作用的抑制。我们发现 LRR 结构域在 D491V 介导的全长 (FL) Pm21 自激活中起着至关重要的作用。 CC 结构域中的一些突变导致Pm21对白粉病的抗性丧失，从而损害了 CC 诱导细胞死亡的活性。两种突变（R73Q 和 E80K）干扰 D491V 介导的 Pm21 自动激活，但不影响 eCC 结构域的细胞死亡诱导活性。值得注意的是，一些在 CC 结构域中携带突变的易感突变体仍然表现出细胞死亡诱导活性。综上所述，这些结果表明 Pm21 的 CC 结构域参与细胞死亡信号传导和抗病信号传导，它们可能彼此独立。

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Pm21 CC domain activity modulated by intramolecular interactions is implicated in cell death and disease resistance.

Nucleotide‐binding (NB) leucine‐rich repeat (LRR) receptors (NLRs) provide resistance against several plant pathogens. We previously cloned the wheat powdery mildew resistance gene Pm21, which encodes a coiled‐coil (CC) NLR that confers broad‐spectrum resistance against Blumeria graminis f. sp. tritici. Here, we report comprehensive biochemical and functional analyses of Pm21 CC domain in Nicotiana benthamiana. Transient overexpression assay suggested that only the extended CC (eCC, amino acid residues 1–159) domain has cell‐death‐inducing activity, whereas the CC‐containing truncations, including CC‐NB and CC‐NB‐LRR, do not induce cell‐death responses. Coimmunoprecipitation (Co‐IP) assay showed that the eCC domain self‐associates and interacts with the NB and LRR domains in planta. These results imply that the activity of the eCC domain is inhibited by the intramolecular interactions of different domains in the absence of pathogens. We found that the LRR domain plays a crucial role in D491V‐mediated full‐length (FL) Pm21 autoactivation. Some mutations in the CC domain leading to the loss of Pm21 resistance to powdery mildew impaired the CC activity of cell‐death induction. Two mutations (R73Q and E80K) interfered with D491V‐mediated Pm21 autoactivation without affecting the cell‐death‐inducing activity of the eCC domain. Notably, some susceptible mutants harbouring mutations in the CC domain still exhibited cell‐death‐inducing activity. Taken together, these results implicate the CC domain of Pm21 in cell‐death signalling and disease‐resistance signalling, which are potentially independent of each other.

更新日期：2020-05-18

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