Pittman Jon K, Edmond Clare, Sunderland Paul A, Bray Clifford M
Faculty of Life Sciences, University of Manchester, Manchester, M13 9PT, United Kingdom.
Faculty of Life Sciences, University of Manchester, Manchester, M13 9PT, United Kingdom.
J Biol Chem. 2009 Jan 2;284(1):525-533. doi: 10.1074/jbc.M807173200. Epub 2008 Nov 10.
The CrCAX1 gene encoding a Ca2+/H+ and Na+/H+ exchanger was cloned and characterized from the unicellular green alga Chlamydomonas reinhardtii to begin to understand the mechanisms of cation homeostasis in this model organism. CrCAX1 was more closely related to fungal cation exchanger (CAX) genes than those from higher plants but has structural characteristics similar to plant Ca2+/H+ exchangers including a long N-terminal tail. When CrCAX1-GFP was expressed in Saccharomyces cerevisiae, it localized at the vacuole. CrCAX1 could suppress the Ca2+-hypersensitive phenotype of a yeast mutant and mediated proton gradient-dependent Ca2+/H+ exchange activity in vacuolar membrane vesicles. Ca2+ transport activity was increased following N-terminal truncation of CrCAX1, suggesting the existence of an N-terminal auto-regulatory mechanism. CrCAX1 could also provide tolerance to Na+ stress when expressed in yeast or Arabidopsis thaliana because of Na+/H+ exchange activity. This Na+/H+ exchange activity was not regulated by the N terminus of the CrCAX1 protein. A subtle tolerance by CrCAX1 in yeast to Co2+ stress was also observed. CrCAX1 was transcriptionally regulated in Chlamydomonas cells grown in elevated Ca2+ or Na+. This study has thus uncovered a novel eukaryotic proton-coupled transporter, CrCAX1, that can transport both monovalent and divalent cations and that appears to play a role in cellular cation homeostasis by the transport of Ca2+ and Na+ into the vacuole.
从单细胞绿藻莱茵衣藻中克隆并鉴定了编码Ca2+/H+和Na+/H+交换体的CrCAX1基因,以开始了解这种模式生物中阳离子稳态的机制。CrCAX1与真菌阳离子交换体(CAX)基因的亲缘关系比与高等植物的CAX基因更近,但具有与植物Ca2+/H+交换体相似的结构特征,包括一个长的N端尾巴。当CrCAX1-GFP在酿酒酵母中表达时,它定位于液泡。CrCAX1可以抑制酵母突变体的Ca2+超敏表型,并介导液泡膜囊泡中质子梯度依赖性的Ca2+/H+交换活性。CrCAX1的N端截短后,Ca2+转运活性增加,表明存在N端自调节机制。由于具有Na+/H+交换活性,CrCAX1在酵母或拟南芥中表达时也能提供对Na+胁迫的耐受性。这种Na+/H+交换活性不受CrCAX1蛋白N端的调节。还观察到CrCAX1在酵母中对Co2+胁迫有轻微的耐受性。在高Ca2+或Na+条件下生长的衣藻细胞中,CrCAX1的转录受到调控。因此,本研究发现了一种新型的真核质子偶联转运体CrCAX1,它可以转运单价和二价阳离子,并且似乎通过将Ca2+和Na+转运到液泡中而在细胞阳离子稳态中发挥作用。