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含有棉花LIM结构域的蛋白GhPLIM1在花药中特异性表达,并参与调节F-肌动蛋白。

Cotton LIM domain-containing protein GhPLIM1 is specifically expressed in anthers and participates in modulating F-actin.

作者信息

Li L, Li Y, Wang N-N, Li Y, Lu R, Li X-B

机构信息

Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Sciences, Central China Normal University, Wuhan, China.

出版信息

Plant Biol (Stuttg). 2015 Mar;17(2):528-34. doi: 10.1111/plb.12243. Epub 2014 Oct 7.

Abstract

As one form of actin binding protein (ABP), LIM domain protein can trigger the formation of actin bundles during plant growth and development. In this study, a cDNA (designated GhPLIM1) encoding a LIM domain protein with 216 amino acid residues was identified from a cotton flower cDNA library. Quantitative RT-PCR indicated that GhPLIM1 is specifically expressed in cotton anthers, and its expression levels are regulated during anther development of cotton. GhPLIM1:eGFP transformed cotton cells display a distributed network of eGFP fluorescence, suggesting that GhPLIM1 protein is mainly localised to the cell cytoskeleton. In vitro high-speed co-sedimentation and low co-sedimentation assays indicate that GhPLIM1 protein not only directly binds actin filaments but also bundles F-actin. Further biochemical experiments verified that GhPLIM1 protein can protect F-actin against depolymerisation by Lat B. Thus, our data demonstrate that GhPLIM1 functions as an actin binding protein (ABP) in modulating actin filaments in vitro, suggesting that GhPLIM1 may be involved in regulating the actin cytoskeleton required for pollen development in cotton.

摘要

作为肌动蛋白结合蛋白(ABP)的一种形式,LIM结构域蛋白在植物生长发育过程中可触发肌动蛋白束的形成。在本研究中,从棉花花cDNA文库中鉴定出一个编码含216个氨基酸残基的LIM结构域蛋白的cDNA(命名为GhPLIM1)。定量RT-PCR表明,GhPLIM1在棉花花药中特异性表达,且其表达水平在棉花花药发育过程中受到调控。GhPLIM1:eGFP转化的棉花细胞显示出eGFP荧光的分布式网络,表明GhPLIM1蛋白主要定位于细胞骨架。体外高速共沉降和低速共沉降试验表明,GhPLIM1蛋白不仅直接结合肌动蛋白丝,还能使F-肌动蛋白成束。进一步的生化实验证实,GhPLIM1蛋白可保护F-肌动蛋白不被Lat B解聚。因此,我们的数据表明,GhPLIM1在体外作为一种肌动蛋白结合蛋白(ABP)发挥作用,调节肌动蛋白丝,提示GhPLIM1可能参与调控棉花花粉发育所需的肌动蛋白细胞骨架。

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