Analysis of highly ordered architecture of actin cytoskeleton in plant cells.

植物细胞中肌动蛋白细胞骨架的高度有序结构分析。

• 批准号: 14540599
• 负责人: YOKOTA Etsuo
• 金额: $ 2.24万
• 依托单位: University of Hyogo (2004)Himeji Institute of Technology (2002-2003)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14540599/
• 关键词: Actin binding protein; Villin; Actin; Pollen; Calcium ion; Lily; Profilin; Tobacco BY-2 cell; カルモジュリン

Plant villins, P-115-ABP and P-135-ABP, containing in a crude extract prepared from lily pollen. bound to a DNase I affinity column in the presence of Ca^<2+>, and were dissociated and eluted from the column with EGTA. Purified P-135-ABP accelerated the polymerization of G-actin in the presence of Ca^<2+>. These results indicated that P-135-ABP can form a complex with G-actin in the presence of Ca^<2+> and consequently acts as a nucleus or seed for polymerization of actin filaments. Furthermore, P-135-ABP caped an actin filament end, and severed or depolymerized filaments in the presence of Ca^<2+>. P-135-ABP has been shown to arrange actin filaments into bundles. Hence, like animal villin, P-135-ABP exerts various effects on actin in a Ca^<2+>-dependent manner. Based on an analysis of the primary structure of P-115-ABP, it is assumed that P-115-ABP also has similar activities to those of P-135-ABP. Hence, plant villin in pollen tubes is responsible for bundling actin filaments in shank region, where low concentrations of Ca^<2+> are present, while is involved in depolymerizing or severing actin filaments in apical regions, where relatively high concentrations of Ca^<2+> concentrations are present. In the presence of profilin, a low molecular weight G-actin binding protein, the acceleration of actin polymerization by P-135-ABP was not found. It is indicated that almost of G-actin form complex with profilin present at similar concentration to that of actin in the germinating pollen. Hence, it is reasonable to consider that actin-nucleation activity of plant villin is not exerted in pollen.Villin is widely distributed in plant cells, such as tobacco cultured cells BY-2. Interestingly, villin is co-localized with not only actin-filament bundles, but also certain organelles considered to be plastides. Therefore, it is possible that plant villin is one of factor responsible in the positioning of the organelles in cells.

从百合花粉中提取的粗提物中含有植物绒毛素 P-115-ABP 和 P-135-ABP。在Ca 2+ 存在下结合至DNase I亲和柱，并用EGTA解离并从柱上洗脱。纯化的P-135-ABP在Ca 2+ 存在下加速G-肌动蛋白的聚合。这些结果表明P-135-ABP可以在Ca 2+ 存在下与G-肌动蛋白形成复合物，并因此充当肌动蛋白丝聚合的核或种子。此外，P-135-ABP覆盖肌动蛋白丝末端，并在Ca 2+ 存在下切断或解聚丝。 P-135-ABP 已被证明可以将肌动蛋白丝排列成束。因此，像动物绒毛一样，P-135-ABP以Ca 2+ 依赖性方式对肌动蛋白发挥多种作用。基于对 P-115-ABP 一级结构的分析，推测 P-115-ABP 也具有与 P-135-ABP 类似的活性。因此，花粉管中的植物绒毛负责在柄区域中捆绑肌动蛋白丝，其中存在低浓度的Ca 2+ ，同时参与解聚或切断顶端区域中的肌动蛋白丝，其中存在相对高浓度的Ca 2+ 。存在<2+>浓度。在 profilin（一种低分子量 G 肌动蛋白结合蛋白）存在的情况下，未发现 P-135-ABP 加速肌动蛋白聚合。这表明几乎大部分G-肌动蛋白与Profilin形成复合物，其浓度与萌发花粉中肌动蛋白的浓度相似。因此，有理由认为植物绒毛蛋白的肌动蛋白成核活性在花粉中不发挥作用。绒毛蛋白广泛分布于植物细胞中，如烟草培养细胞BY-2。有趣的是，绒毛不仅与肌动蛋白丝束共定位，还与某些被认为是质体的细胞器共定位。因此，植物绒毛可能是负责细胞器在细胞中定位的因素之一。

项目成果

E.Yokota: "Plant 115-kDa-actin-filament-bundling protein, P-115-ABP, is a homologue of plant villin and is widely distributed in cells."Plant Cell Physiol.. 44. 1088-1099 (2003)

E.Yokota：“植物 115-kDa-肌动蛋白丝束蛋白，P-115-ABP，是植物绒毛蛋白的同源物，广泛分布在细胞中。”Plant Cell Physiol.. 44. 1088-1099 (2003)

T.Nishimura: "A semi-dominant mutation in the ACT2 gene affects the root hair development in Arabidopsis."Plant Cell Physiol. 44. 1131-1140 (2003)

T.Nishimura：“ACT2 基因的半显性突变影响拟南芥根毛的发育。”植物细胞生理学。

M.Tominaga: "Higher plant myosin XI moves processively on actin with 35 nm steps at high velocity."EMBO J.. 22. 1263-1272 (2003)

M.Tominaga：“高等植物肌球蛋白 XI 在肌动蛋白上以 35 nm 的步长高速连续移动。”EMBO J.. 22. 1263-1272 (2003)

Plant 115-kDa actin-filament bundling protein, P-115-ABP, is a homologue of plant villin and is widely distributed in cells.

• DOI: 10.1093/pcp/pcg132
• 发表时间: 2003-10
• 期刊: Plant & cell physiology
• 影响因子: 4.9
• 作者: E. Yokota;L. Vidali;M. Tominaga;H. Tahara;H. Orii;Yosuke Morizane;P. K. Hepler;T. Shimmen

E.Yokota: "Possible involvement of energy metabolism in the change of cytoplasm organization induced by a protein phosphatase inhibitor, calyculin A, in root hair cells of Limnobium."Protoplasma. 221. 217-226 (2003)

E.Yokota：“能量代谢可能参与由蛋白磷酸酶抑制剂花萼蛋白 A 在 Limnobium 根毛细胞中诱导的细胞质组织变化。”原生质体。