Jain Deepti, Roy Nilanjan, Chattopadhyay Debasis
National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, India.
PLoS One. 2009;4(4):e5154. doi: 10.1371/journal.pone.0005154. Epub 2009 Apr 13.
Salt-sensitive yeast mutants were deployed to characterize a gene encoding a C2H2 zinc finger protein (CaZF) that is differentially expressed in a drought-tolerant variety of chickpea (Cicer arietinum) and provides salinity-tolerance in transgenic tobacco. In Saccharomyces cerevisiae most of the cellular responses to hyper-osmotic stress is regulated by two interconnected pathways involving high osmolarity glycerol mitogen-activated protein kinase (Hog1p) and Calcineurin (CAN), a Ca(2+)/calmodulin-regulated protein phosphatase 2B. In this study, we report that heterologous expression of CaZF provides osmotolerance in S. cerevisiae through Hog1p and Calcineurin dependent as well as independent pathways. CaZF partially suppresses salt-hypersensitive phenotypes of hog1, can and hog1can mutants and in conjunction, stimulates HOG and CAN pathway genes with subsequent accumulation of glycerol in absence of Hog1p and CAN. CaZF directly binds to stress response element (STRE) to activate STRE-containing promoter in yeast. Transactivation and salt tolerance assays of CaZF deletion mutants showed that other than the transactivation domain a C-terminal domain composed of acidic and basic amino acids is also required for its function. Altogether, results from this study suggests that CaZF is a potential plant salt-tolerance determinant and also provide evidence that in budding yeast expression of HOG and CAN pathway genes can be stimulated in absence of their regulatory enzymes to provide osmotolerance.
利用盐敏感酵母突变体来表征一个编码C2H2锌指蛋白(CaZF)的基因,该基因在耐旱型鹰嘴豆(Cicer arietinum)中差异表达,并在转基因烟草中赋予耐盐性。在酿酒酵母中,细胞对高渗胁迫的大多数反应由两条相互关联的途径调节,这两条途径涉及高渗甘油丝裂原活化蛋白激酶(Hog1p)和钙调神经磷酸酶(CAN),一种Ca(2+)/钙调蛋白调节的蛋白磷酸酶2B。在本研究中,我们报告CaZF的异源表达通过依赖和不依赖Hog1p及钙调神经磷酸酶的途径赋予酿酒酵母渗透耐受性。CaZF部分抑制hog1、can和hog1can突变体的盐超敏表型,并在没有Hog1p和CAN的情况下,协同刺激HOG和CAN途径基因,随后积累甘油。CaZF直接结合应激反应元件(STRE)以激活酵母中含STRE的启动子。CaZF缺失突变体的反式激活和耐盐性分析表明,除反式激活域外,由酸性和碱性氨基酸组成的C末端结构域对其功能也是必需的。总之,本研究结果表明CaZF是一种潜在的植物耐盐决定因素,也提供了证据表明在出芽酵母中,在没有其调节酶的情况下,HOG和CAN途径基因的表达可以被刺激以提供渗透耐受性。