Noël Laurent D, Cagna Giuseppe, Stuttmann Johannes, Wirthmüller Lennart, Betsuyaku Shigeyuki, Witte Claus-Peter, Bhat Riyaz, Pochon Nathalie, Colby Thomas, Parker Jane E
Institut de Biologie Environementale et Biotechnologie, Unité Mixte de Recherche 6191, Centre National de la Recherche Scientifique, Université de la Méditerranée Aix-Marseille II, Saint Paul-lez-Durance Cedex, France.
Plant Cell. 2007 Dec;19(12):4061-76. doi: 10.1105/tpc.107.051896. Epub 2007 Dec 7.
The conserved eukaryotic protein SGT1 (for Suppressor of G2 allele of skp1) has characteristics of an HSP90 (for heat shock protein 90 kD) cochaperone and in plants regulates hormone responses and Resistance gene-triggered immunity. We affinity-purified SGT1-interacting proteins from Arabidopsis thaliana leaf extracts and identified by mass spectrometry cytosolic heat shock cognate 70 (HSC70) chaperones as the major stable SGT1 interactors. Arabidopsis SGT1a and SGT1b proteins associate with HSC70 in vivo and distribute with HSC70 in the cytosol and nucleus. An intact C-terminal SGT1-specific (SGS) domain that is required for all known SGT1b functions in immunity and development is needed for HSC70 interaction and for the nuclear accumulation of SGT1b. Interaction assays of transiently expressed proteins or their domains in Nicotiana benthamiana point to a role of SGT1 as a HSC70 cofactor. Expression of two HSC70 isoforms is upregulated by pathogen challenge, and while loss of function of individual cytosolic HSC70 genes has no defense phenotype, HSC70-1 overexpression disables resistance to virulent and avirulent pathogens. Moreover, mutations in SGT1b lead to a similar degree of heat shock tolerance as deregulation of HSC70-1. We conclude that an HSC70-SGT1 chaperone complex is important for multiple plant environmental responses and that the evolutionarily conserved SGS domain of SGT1 is a key determinant of the HSC70-SGT1 association.
保守的真核蛋白SGT1(skp1的G2等位基因的抑制因子)具有热休克蛋白90(HSP90,90kD热休克蛋白)伴侣蛋白的特征,在植物中调节激素反应和抗性基因触发的免疫反应。我们从拟南芥叶片提取物中亲和纯化了与SGT1相互作用的蛋白,并通过质谱鉴定出胞质热休克同源蛋白70(HSC70)伴侣蛋白是主要的稳定SGT1相互作用蛋白。拟南芥SGT1a和SGT1b蛋白在体内与HSC70结合,并与HSC70一起分布在细胞质和细胞核中。HSC70相互作用以及SGT1b的核积累需要完整的C末端SGT1特异性(SGS)结构域,该结构域是SGT1b在免疫和发育中所有已知功能所必需的。在本氏烟草中对瞬时表达的蛋白或其结构域进行的相互作用分析表明,SGT1作为HSC70辅因子发挥作用。病原体攻击会上调两种HSC70亚型的表达,虽然单个胞质HSC70基因功能丧失没有防御表型,但HSC70-1过表达会使植物对毒性和无毒病原体的抗性丧失。此外,SGT1b中的突变导致的热休克耐受性程度与HSC70-1失调相似。我们得出结论,HSC70-SGT1伴侣蛋白复合物对多种植物环境反应很重要,并且SGT1进化上保守的SGS结构域是HSC70-SGT1关联的关键决定因素。