Yamada Nana, Theerawitaya Cattarin, Kageyama Hakuto, Cha-Um Suriyan, Takabe Teruhiro
Plant Physiology and Biochemistry Laboratory, BIOTEC, Pathumthani, 12120, Thailand.
Graduate School of Environmental and Human Sciences, Meijo University, Nagoya, 468-8502, Japan.
Protoplasma. 2015 Nov;252(6):1519-27. doi: 10.1007/s00709-015-0781-x. Epub 2015 Mar 6.
The cytoplasmic free Ca(2+) could play an important role for salt tolerance in rice root (Oryza sativa L.). Here, we compared the expression profiles of two putative developmentally regulated plasma membrane polypeptides (DREPP1 and DREPP2) in rice roots of salt-tolerant cv. Pokkali and salt-sensitive cv. IR29. The messenger RNA (mRNA) for OsDREPP1 could be detected in all parts of root and did not change upon salt stress, whereas the mRNA for OsDREPP2 was detected only in root tips. The transcript level of OsDREPP2 first disappeared upon salt stress, then recovered in Pokkali, but not recovered in IR29. The gene-encoding OsDREPP2 was cloned from cv. Pokkali and expressed in Escherichia coli, and its biochemical properties were studied. It was found that OsDREPP2 is a Ca(2+)-binding protein and binds also to calmodulin (CaM) as well as microtubules. The mutation of Trp4 and Phe16 in OsDREPP2 to Ala decreased the binding of DREPP2 to Ca(2+)/CaM complex, indicating the N-terminal basic domain is involved for the binding. The binding of OsDREPP2 to microtubules was inhibited by Ca(2+)/CaM complex, while the binding of double-mutant OsDREPP2 protein to microtubules was not inhibited by Ca(2+)/CaM complex. We propose that CaM inhibits the binding of DREPP2 to cortical microtubules, causes the inhibition of microtubule depolymerization, and enhances the cell elongation.
细胞质游离Ca(2+)可能在水稻根(Oryza sativa L.)耐盐性中发挥重要作用。在此,我们比较了耐盐品种Pokkali和盐敏感品种IR29的水稻根中两种假定的发育调控质膜多肽(DREPP1和DREPP2)的表达谱。OsDREPP1的信使核糖核酸(mRNA)在根的所有部位均可检测到,且在盐胁迫下不发生变化,而OsDREPP2的mRNA仅在根尖中检测到。OsDREPP2的转录水平在盐胁迫下首先消失,然后在Pokkali中恢复,但在IR29中未恢复。从Pokkali品种中克隆了编码OsDREPP2的基因并在大肠杆菌中表达,对其生化特性进行了研究。发现OsDREPP2是一种Ca(2+)结合蛋白,也与钙调蛋白(CaM)以及微管结合。OsDREPP2中Trp4和Phe16突变为Ala降低了DREPP2与Ca(2+)/CaM复合物的结合,表明N端碱性结构域参与该结合。Ca(2+)/CaM复合物抑制OsDREPP2与微管的结合,而双突变OsDREPP2蛋白与微管的结合不受Ca(2+)/CaM复合物的抑制。我们提出,CaM抑制DREPP2与皮层微管的结合,导致微管解聚的抑制,并促进细胞伸长。