Kudla J, Xu Q, Harter K, Gruissem W, Luan S
Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA.
Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4718-23. doi: 10.1073/pnas.96.8.4718.
An important effector of Ca2+ signaling in animals and yeast is the Ca2+/calmodulin-dependent protein phosphatase calcineurin. However, the biochemical identity of plant calcineurin remained elusive. Here we report the molecular characterization of AtCBL (Arabidopsis thaliana calcineurin B-like protein) from Arabidopsis. The protein is most similar to mammalian calcineurin B, the regulatory subunit of the phosphatase. AtCBL also shows significant similarity with another Ca2+-binding protein, the neuronal calcium sensor in animals. It contains typical EF-hand motifs with Ca2+-binding capability, as confirmed by in vitro Ca2+-binding assays, and it interacts in vivo with rat calcineurin A in the yeast two-hybrid system. Interaction of AtCBL1 and rat calcineurin A complemented the salt-sensitive phenotype in a yeast calcineurin B mutant. Cloning of cDNAs revealed that AtCBL proteins are encoded by a family of at least six genes in Arabidopsis. Genes for three isoforms were identified in this study. AtCBL1 mRNA was preferentially expressed in stems and roots and its mRNA levels strongly increased in response to specific stress signals such as drought, cold, and wounding. In contrast, AtCBL2 and AtCBL3 are constitutively expressed under all conditions investigated. Our data suggest that AtCBL1 may act as a regulatory subunit of a plant calcineurin-like activity mediating calcium signaling under certain stress conditions.
动物和酵母中Ca2+信号的一个重要效应器是Ca2+/钙调蛋白依赖性蛋白磷酸酶钙调神经磷酸酶。然而,植物钙调神经磷酸酶的生化特性仍然难以捉摸。在此,我们报道了来自拟南芥的AtCBL(拟南芥钙调神经磷酸酶B样蛋白)的分子特征。该蛋白与哺乳动物钙调神经磷酸酶B(磷酸酶的调节亚基)最为相似。AtCBL也与另一种Ca2+结合蛋白——动物中的神经元钙传感器有显著相似性。通过体外Ca2+结合试验证实,它含有具有Ca2+结合能力的典型EF手基序,并且在酵母双杂交系统中它在体内与大鼠钙调神经磷酸酶A相互作用。AtCBL1与大鼠钙调神经磷酸酶A的相互作用弥补了酵母钙调神经磷酸酶B突变体中的盐敏感表型。cDNA的克隆表明,AtCBL蛋白由拟南芥中至少六个基因组成的家族编码。本研究鉴定了三种同工型的基因。AtCBL1 mRNA在茎和根中优先表达,其mRNA水平在响应干旱、寒冷和创伤等特定应激信号时强烈增加。相比之下,AtCBL2和AtCBL3在所有研究条件下均组成型表达。我们的数据表明,AtCBL1可能作为植物钙调神经磷酸酶样活性的调节亚基,在某些应激条件下介导钙信号传导。