Ma Lian, Xie Bo, Hong Zonglie, Verma Desh Pal S, Zhang Zhongming
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
Plant Physiol. 2008 Sep;148(1):223-34. doi: 10.1104/pp.108.120527. Epub 2008 Jul 11.
Building a cell plate during cytokinesis in plant cells requires the participation of a number of proteins in a multistep process. We previously identified phragmoplastin as a cell plate-specific protein involved in creating a tubulovesicular network at the cell plate. We report here the identification and characterization of a phragmoplastin-interacting protein, PHIP1, in Arabidopsis (Arabidopsis thaliana). It contains multiple functional motifs, including a lysine-rich domain, two RNA recognition motifs, and three CCHC-type zinc fingers. Polypeptides with similar motif structures were found only in plant protein databases, but not in the sequenced prokaryotic, fungal, and animal genomes, suggesting that PHIP1 represents a plant-specific RNA-binding protein. In addition to phragmoplastin, two Arabidopsis small GTP-binding proteins, Rop1 and Ran2, are also found to interact with PHIP1. The zinc fingers of PHIP1 were not required for its interaction with Rop1 and phragmoplastin, but they may participate in its binding with the Ran2 mRNA. Immunofluorescence, in situ RNA hybridization, and green fluorescent protein tagging experiments showed the association of PHIP1 with the forming cell plate during cytokinesis. Taken together, our data suggest that PHIP1 is a novel RNA-binding protein and may play a unique role in the polarized mRNA transport to the vicinity of the cell plate.
在植物细胞胞质分裂过程中构建细胞板需要多种蛋白质参与一个多步骤过程。我们之前鉴定出成膜体蛋白是一种细胞板特异性蛋白,参与在细胞板处形成微管泡网络。我们在此报告在拟南芥中一种与成膜体蛋白相互作用的蛋白PHIP1的鉴定和特性。它包含多个功能基序,包括一个富含赖氨酸的结构域、两个RNA识别基序和三个CCHC型锌指结构。仅在植物蛋白质数据库中发现具有相似基序结构的多肽,而在已测序的原核生物、真菌和动物基因组中未发现,这表明PHIP1代表一种植物特异性RNA结合蛋白。除了成膜体蛋白外,还发现两种拟南芥小GTP结合蛋白Rop1和Ran2与PHIP1相互作用。PHIP1的锌指结构对于其与Rop1和成膜体蛋白的相互作用不是必需的,但它们可能参与其与Ran2 mRNA的结合。免疫荧光、原位RNA杂交和绿色荧光蛋白标记实验表明在胞质分裂期间PHIP1与正在形成的细胞板相关联。综上所述,我们的数据表明PHIP1是一种新型RNA结合蛋白,可能在将极化的mRNA转运到细胞板附近发挥独特作用。