Borsics Tamás, Webb David, Andeme-Ondzighi Christine, Staehelin L Andrew, Christopher David A
Department of Molecular Biosciences and Bioengineering, University of Hawaii, 1955 East-West Road, Agsciences 218, Honolulu, HI 96822, USA.
Planta. 2007 Feb;225(3):563-73. doi: 10.1007/s00425-006-0372-3. Epub 2006 Aug 31.
Cyclic nucleotide gated channels (CNGCs) that are regulated by calmodulin (CaM) have been shown to play essential roles in signal transduction, metabolism, and growth in animals. By contrast, very little is known about the subcellular location and the function of these channels in plants. Here we report on the effects of antisense suppression of the expression of AtCNGC10, a putative K+ channel, and the immunolocalization of the protein using an AtCNGC10-specific antiserum. In Arabidopsis thaliana leaves, AtCNGC10 was localized to the plasma membrane of mesophyll and parenchyma cells. Antisense AtCNGC10 plants had 40% of the AtCNGC10 mRNA levels and virtually undetectable protein levels relative to wild type plants. Antisense expression of AtCNGC10 did not affect the mRNA levels of AtCNGC13, the most closely related CNGC family member in the genome. Relative to wild type Columbia, antisense AtCNGC10 plants flowered 10 days earlier, and had a 25% reduction in leaf surface area, thickness and palisade parenchyma cell length. Their roots responded more slowly to gravitropic changes and the chloroplasts accumulated more starch. We propose that AtCNGC10, through interactions with CaM and cGMP, modulates cellular K+ balance across the plasma membrane, and that perturbations of this K+ gradient affect numerous growth and developmental processes.
受钙调蛋白(CaM)调节的环核苷酸门控通道(CNGCs)已被证明在动物的信号转导、代谢和生长中发挥重要作用。相比之下,关于这些通道在植物中的亚细胞定位和功能却知之甚少。在此,我们报告了对拟钾离子通道AtCNGC10表达进行反义抑制的效果,以及使用AtCNGC10特异性抗血清对该蛋白进行免疫定位的情况。在拟南芥叶片中,AtCNGC10定位于叶肉细胞和薄壁细胞的质膜。与野生型植株相比,反义AtCNGC10植株的AtCNGC10 mRNA水平仅为40%,且蛋白水平几乎检测不到。AtCNGC10的反义表达不影响基因组中与之关系最密切的CNGC家族成员AtCNGC13的mRNA水平。相对于野生型哥伦比亚植株,反义AtCNGC10植株提前10天开花,叶片表面积、厚度和栅栏薄壁细胞长度减少25%。它们的根对重力变化的反应更慢,叶绿体积累了更多淀粉。我们推测,AtCNGC10通过与CaM和cGMP相互作用,调节质膜上的细胞钾离子平衡,而这种钾离子梯度的扰动会影响众多生长和发育过程。