Lechner Esther, Xie Daoxin, Grava Sandrine, Pigaglio Emmanuelle, Planchais Severine, Murray James A H, Parmentier Yves, Mutterer Jerome, Dubreucq Bertrand, Shen Wen-Hui, Genschik Pascal
Institut de Biologie Moléculaire des Plantes du CNRS, 12, rue du Général Zimmer, 67084 Strasbourg Cédex, France.
J Biol Chem. 2002 Dec 20;277(51):50069-80. doi: 10.1074/jbc.M204254200. Epub 2002 Oct 14.
Recently in yeast and animal cells, one particular class of ubiquitin ligase (E3), called the SCF, was demonstrated to regulate diverse processes including cell cycle and development. In plants SCF-dependent proteolysis is also involved in different developmental and hormonal regulations. To further investigate the function of SCF, we characterized at the molecular level the Arabidopsis RING-H2 finger protein AtRbx1. We demonstrated that the plant gene is able to functionally complement a yeast knockout mutant strain and showed that AtRbx1 protein interacts physically with at least two members of the Arabidopsis cullin family (AtCul1 and AtCul4). AtRbx1 also associates with AtCul1 and the Arabidopsis SKP1-related proteins in planta, indicating that it is part of plant SCF complexes. AtRbx1 mRNAs accumulate in various tissues of the plant, but at higher levels in tissues containing actively dividing cells. Finally to study the function of the gene in planta, we either overexpressed AtRbx1 or reduced its expression by a dsRNA strategy. Down-regulation of AtRbx1 impaired seedling growth and development, indicating that the gene is essential in plants. Furthermore, the AtRbx1-silenced plants showed a reduced level of AtCul1 protein, but accumulated higher level of cyclin D3.
最近在酵母和动物细胞中,一种特定类型的泛素连接酶(E3),即SCF,被证明可调节包括细胞周期和发育在内的多种过程。在植物中,SCF依赖的蛋白水解作用也参与不同的发育和激素调节。为了进一步研究SCF的功能,我们在分子水平上对拟南芥RING-H2指蛋白AtRbx1进行了表征。我们证明该植物基因能够在功能上互补酵母敲除突变株,并表明AtRbx1蛋白与拟南芥cullin家族的至少两个成员(AtCul1和AtCul4)发生物理相互作用。AtRbx1在植物中还与AtCul1和拟南芥SKP1相关蛋白结合,表明它是植物SCF复合物的一部分。AtRbx1 mRNA在植物的各种组织中积累,但在含有活跃分裂细胞的组织中水平更高。最后,为了研究该基因在植物中的功能,我们要么过表达AtRbx1,要么通过dsRNA策略降低其表达。AtRbx1的下调损害了幼苗的生长和发育,表明该基因在植物中至关重要。此外,AtRbx1沉默的植物显示AtCul1蛋白水平降低,但细胞周期蛋白D3水平升高。