Garciadeblás Blanca, Haro Rosario, Benito Begoña
Departamento de Biotecnología, Escuela Técnica Superior de Ingenieros Agrónomos, Universidad Politécnica de Madrid, 28040 Madrid, Spain.
Plant Mol Biol. 2007 Mar;63(4):479-90. doi: 10.1007/s11103-006-9102-2.
Two cDNAs isolated from Cymodocea nodosa, CnSOS1A, and CnSOSIB encode proteins with high-sequence similarities to SOS1 plant transporters. CnSOS1A expressed in a yeast Na+-efflux mutant under the control of a constitutive expression promoter mimicked AtSOS1 from Arabidopsis; the wild type cDNA did not improve the growth of the recipient strain in the presence of Na+, but a cDNA mutant that expresses a truncated protein suppressed the defect of the yeast mutant. In similar experiments, CnSOS1B was not effective. Conditional expression, under the control of an arabinose responsive promoter, of the CnSOSIA and CnSOS1B cDNAs in an Escherichia coli mutant defective in Na+ efflux was toxic, and functional analyses were inconclusive. The same constructs transformed into an E. coli K+-uptake mutant revealed that CnSOS1A was also toxic, but that it slightly suppressed defective growth at low K+. Truncation in the C-terminal hydrophilic tail of CnSOS1A relieved the toxicity and proved that CnSOS1A was an excellent low-affinity K+ and Rb+ transporter. CnSOS1B mediated a transient, extremely rapid K+ or Rb+ influx. Similar tests with AtSOS1 revealed that it was not toxic and that the whole protein exhibited excellent K+ and Rb+ uptake characteristics in bacteria.
从波喜荡草中分离出的两个cDNA,即CnSOS1A和CnSOS1B,编码的蛋白质与SOS1植物转运蛋白具有高度的序列相似性。在组成型表达启动子的控制下,在酵母Na⁺外流突变体中表达的CnSOS1A模拟了拟南芥中的AtSOS1;野生型cDNA在存在Na⁺的情况下并不能改善受体菌株的生长,但表达截短蛋白的cDNA突变体抑制了酵母突变体的缺陷。在类似的实验中,CnSOS1B无效。在阿拉伯糖响应启动子的控制下,在Na⁺外流缺陷的大肠杆菌突变体中条件性表达CnSOS1A和CnSOS1B的cDNA具有毒性,功能分析尚无定论。将相同的构建体转化到大肠杆菌K⁺摄取突变体中发现,CnSOS1A也具有毒性,但它在低钾条件下略微抑制了缺陷生长。CnSOS1A C末端亲水尾巴的截短消除了毒性,并证明CnSOS1A是一种出色的低亲和力K⁺和Rb⁺转运蛋白。CnSOS1B介导了短暂、极其快速的K⁺或Rb⁺内流。对AtSOS1进行的类似测试表明,它没有毒性,并且整个蛋白在细菌中表现出出色的K⁺和Rb⁺摄取特性。