Mizoguchi T, Irie K, Hirayama T, Hayashida N, Yamaguchi-Shinozaki K, Matsumoto K, Shinozaki K
Laboratory of Plant Molecular Biology, Institute of Physical and Chemical Research (RIKEN), Tsukuba Life Science Center, Ibaraki, Japan.
Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):765-9. doi: 10.1073/pnas.93.2.765.
We describe here the cloning and characterization of a cDNA encoding a protein kinase that has high sequence homology to members of the mitogen-activated protein kinase (MAPK) kinase kinase (MAPKKK or MEKK) family; this cDNA is named cATMEKKI (Arabidopsis thaliana MAP kinase or ERK kinase kinase 1). The catalytic domain of the putative ATMEKK1 protein shows approximately 40% identity with the amino acid sequences of the catalytic domains of MAPKKKs (such as Byr2 from Schizosaccharomyces pombe, Ste11 from Saccharomyces cerevisiae, Bck1 from S. cerevisiae, MEKK from mouse, and NPK1 from tobacco). In yeast cells that overexpress ATMEKK1, the protein kinase replaces Ste11 in responding to mating pheromone. In this study, the expression of three protein kinases was examined by Northern blot analyses: ATMEKK1 (structurally related to MAPKKK), ATMPK3 (structurally related to MAPK), and ATPK19 (structurally related to ribosomal S6 kinase). The mRNA levels of these three protein kinases increased markedly and simultaneously in response to touch, cold, and salinity stress. These results suggest that MAP kinase cascades, which are thought to respond to a variety of extracellular signals, are regulated not only at the posttranslational level but also at the transcriptional level in plants and that MAP kinase cascades in plants may function in transducing signals in the presence of environmental stress.
我们在此描述了一种编码蛋白激酶的cDNA的克隆与特性分析,该蛋白激酶与促分裂原活化蛋白激酶(MAPK)激酶激酶(MAPKKK或MEKK)家族成员具有高度序列同源性;此cDNA被命名为cATMEKKI(拟南芥MAP激酶或ERK激酶激酶1)。推测的ATMEKK1蛋白的催化结构域与MAPKKKs(如粟酒裂殖酵母的Byr2、酿酒酵母的Ste11、酿酒酵母的Bck1、小鼠的MEKK以及烟草的NPK1)催化结构域的氨基酸序列显示出约40%的同一性。在过表达ATMEKK1的酵母细胞中,该蛋白激酶在响应交配信息素时取代了Ste11。在本研究中,通过Northern印迹分析检测了三种蛋白激酶的表达:ATMEKK1(在结构上与MAPKKK相关)、ATMPK3(在结构上与MAPK相关)和ATPK19(在结构上与核糖体S6激酶相关)。这三种蛋白激酶的mRNA水平在受到触摸、低温和盐胁迫时显著且同时升高。这些结果表明,被认为对多种细胞外信号作出反应的MAP激酶级联反应,在植物中不仅在翻译后水平受到调控,而且在转录水平也受到调控,并且植物中的MAP激酶级联反应可能在环境胁迫存在时转导信号。