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Heterodimerization of Arabidopsis calcium/proton exchangers contributes to regulation of guard cell dynamics and plant defense responses

机译:拟南芥钙/质子交换剂的异二聚化有助于调节保卫细胞动力学和植物防御反应

摘要

Arabidopsis thaliana cation exchangers (CAX1 and CAX3) are closely related tonoplast-localized calcium/proton (Ca²⁺/H⁺) antiporters that contribute to cellular Ca²⁺ homeostasis. CAX1 and CAX3 were previously shown to interact in yeast; however, the function of this complex in plants has remained elusive. Here, we demonstrate that expression of CAX1 and CAX3 occurs in guard cells. Additionally, CAX1 and CAX3 are co-expressed in mesophyll tissue in response to wounding or flg22 treatment, due to the induction of CAX3 expression. Having shown that the transporters can be co-expressed in the same cells, we demonstrate that CAX1 and CAX3 can form homomeric and heteromeric complexes in plants. Consistent with the formation of a functional CAX1-CAX3 complex, CAX1 and CAX3 integrated into the yeast genome suppressed a Ca²⁺-hypersensitive phenotype of mutants defective in vacuolar Ca²⁺ transport, and demonstrated enzyme kinetics different from those of either CAX protein expressed by itself. We demonstrate that the interactions between CAX proteins contribute to the functioning of stomata, because stomata were more closed in cax1-1, cax3-1, and cax1-1/cax3-1 loss-of-function mutants due to an inability to buffer Ca²⁺ effectively. We hypothesize that the formation of CAX1-CAX3 complexes may occur in the mesophyll to affect intracellular Ca²⁺ signaling during defense responses.
机译:拟南芥阳离子交换剂(CAX1和CAX3)与原生质体定位的钙/质子(Ca 2+ / H 3)反向转运蛋白密切相关,这些反向转运蛋白有助于细胞内Ca 2 +稳态。先前已证明CAX1和CAX3在酵母中相互作用。然而,这种复合物在植物中的功能仍然难以捉摸。在这里,我们证明了CAX1和CAX3的表达发生在保卫细胞中。另外,由于CAX3表达的诱导,响应于伤口或flg22治疗,CAX1和CAX3在叶肉组织中共表达。已经表明转运蛋白可以在同一细胞中共表达,我们证明了CAX1和CAX3可以在植物中形成同聚和异聚复合物。与功能性CAX1-CAX3复合物的形成一致,整合到酵母基因组中的CAX1和CAX3抑制了液泡Ca2 +转运缺陷型突变体的Ca2 +超敏表型,并证明了与自身表达的任一CAX蛋白不同的酶动力学。 。我们证明了CAX蛋白之间的相互作用有助于气孔的功能,因为由于无法缓冲Ca²,气孔在cax1-1,cax3-1和cax1-1 / cax3-1功能丧失的突变体中更封闭⁺有效。我们假设CAX1-CAX3复合物的形成可能发生在叶肉中,从而在防御反应期间影响细胞内Ca²2信号传导。

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