Matsui M, Stoop C D, von Arnim A G, Wei N, Deng X W
Department of Biology, Yale University, New Haven, CT 06520-8104, USA.
Proc Natl Acad Sci U S A. 1995 May 9;92(10):4239-43. doi: 10.1073/pnas.92.10.4239.
Arabidopsis COP1 acts inside the nucleus to suppress photomorphogenic cellular development, and light inactivation of COP1 may involve a specific control of its nuclear activity in hypocotyls and cotyledons, but not in roots, of developing seedlings. To understand the molecular mechanisms of COP1 action during light-mediated development, we initiated a screen for Arabidopsis cDNAs encoding proteins which interact directly with COP1 in vitro as a step to identify the cellular components involved. We report here the isolation and characterization of a cDNA clone encoding a protein designated CIP1 (COP1-interactive protein 1). CIP1 is predominantly alpha-helical and most likely involved in coiled-coil formation. It interacts specifically with the putative coiled-coil region of COP1 in vitro. Further, CIP1 is encoded by a single gene in Arabidopsis, and its mRNA and protein levels are not regulated by light. Immunofluorescent labeling of CIP1 in Arabidopsis seedling protoplasts demonstrated that CIP1 is part of, or associated with, a cytoskeletal structure in hypocotyl and cotyledon cells, but not in roots. Our results are consistent with a possible role of CIP1 in mediating light control of COP1 nuclear activity by regulating its nucleocytoplasmic partitioning.
拟南芥COP1在细胞核内发挥作用以抑制光形态建成的细胞发育,并且COP1的光失活可能涉及对其在发育幼苗的下胚轴和子叶中而非根中的核活性的特定调控。为了了解COP1在光介导的发育过程中的作用分子机制,我们开展了一项筛选,寻找在体外与COP1直接相互作用的拟南芥cDNA编码蛋白,以此作为鉴定相关细胞成分的第一步。我们在此报告一个编码名为CIP1(COP1相互作用蛋白1)的蛋白的cDNA克隆的分离与鉴定。CIP1主要为α螺旋结构,很可能参与卷曲螺旋的形成。它在体外与COP1假定的卷曲螺旋区域特异性相互作用。此外,CIP1由拟南芥中的单一基因编码,其mRNA和蛋白水平不受光的调控。对拟南芥幼苗原生质体中的CIP1进行免疫荧光标记表明,CIP1是下胚轴和子叶细胞中细胞骨架结构的一部分或与其相关,但在根中并非如此。我们的结果与CIP1通过调节COP1的核质分配来介导对其核活性的光控这一可能作用相一致。