Pseudomonas syringae pv. tomato DC3000 Type III Secretion Effector Polymutants Reveal an Interplay between HopAD1 and AvrPtoB.

Pseudomonas syringae pv. tomato DC3000 Type III Secretion Effector Polymutants Reveal an Interplay between HopAD1 and AvrPtoB.
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DOI:
10.1016/j.chom.2015.05.007
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发表时间:
2015-06-10
影响因子:
30.3
通讯作者:
Collmer A
Collmer A
中科院分区:
医学1区
文献类型:
--
作者:
Wei HL;Chakravarthy S;Mathieu J;Helmann TC;Stodghill P;Swingle B;Martin GB;Collmer A

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细菌病原体丁香假单胞菌 pv。番茄 DC3000 通过注入复杂的 III 型分泌效应蛋白 (T3E) 来抑制两层植物先天免疫系统。除了冗余和相互作用之外,单个 T3E 还可能与多种免疫相关蛋白相互作用,这使得它们的分析具有挑战性。我们构建了缺乏所有 36 个 T3E 的 Pst DC3000 多突变体,并恢复了单个 T3E 或其突变体,以探索 T3E 之间的相互作用。弱表达的 T3E HopAD1 足以在本塞姆氏烟草中引起免疫相关的细胞死亡。 HopAD1 诱导的细胞死亡被天然 AvrPtoB 部分抑制,并被 AvrPtoBM3 完全抑制,AvrPtoBM3 的突变破坏了其 E3 泛素连接酶结构域和两个已知的与免疫相关激酶相互作用的结构域。 AvrPtoBM3 还获得了与免疫激酶 MKK2 相互作用的能力,这是 HopAD1 依赖性细胞死亡所必需的。因此,AvrPtoB 具有替代的竞争机制来抑制效应子触发的植物免疫。这种方法允许对各个 T3E 活动进行反卷积。
The bacterial pathogen Pseudomonas syringae pv. tomato DC3000 suppresses the two-tiered plant innate immune system by injecting a complex repertoire of type III secretion effector (T3E) proteins. Beyond redundancy and interplay, individual T3Es may interact with multiple immunity-associated proteins, rendering their analysis challenging. We constructed a Pst DC3000 polymutant lacking all 36 T3Es and restored individual T3Es or their mutants to explore the interplay among T3Es. The weakly expressed T3E HopAD1 was sufficient to elicit immunity-associated cell death in Nicotiana benthamiana. HopAD1-induced cell death was suppressed partially by native AvrPtoB and completely by AvrPtoBM3, which has mutations disrupting its E3 ubiquitin ligase domain and two known domains for interacting with immunity-associated kinases. AvrPtoBM3 also gained the ability to interact with the immunity-kinase MKK2, which is required for HopAD1-dependent cell death. Thus, AvrPtoB has alternative, competing mechanisms for suppressing effector-triggered plant immunity. This approach allows the deconvolution of individual T3E activities.
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