首页> 美国卫生研究院文献>Proceedings of the National Academy of Sciences of the United States of America >Two interacting coiled-coil proteins WEB1 and PMI2 maintain the chloroplast photorelocation movement velocity in Arabidopsis
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Two interacting coiled-coil proteins WEB1 and PMI2 maintain the chloroplast photorelocation movement velocity in Arabidopsis

机译:两种相互作用的卷曲螺旋蛋白WEB1和PMI2维持拟南芥中叶绿体的光定位运动速度

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摘要

Chloroplasts move toward weak light (accumulation response) and away from strong light (avoidance response). The fast and accurate movement of chloroplasts in response to ambient light conditions is essential for efficient photosynthesis and photodamage prevention in chloroplasts. Here, we report that two Arabidopsis mutants, weak chloroplast movement under blue light 1 (web1) and web2, are defective in both the avoidance and the accumulation responses. Map-based cloning revealed that both genes encode coiled-coil proteins and that WEB2 is identical to the plastid movement impaired 2 (PMI2) gene. The velocities of chloroplast movement in web1 and pmi2 were approximately threefold lower than that in the wild type. Defects in the avoidance response of web1 and pmi2 were suppressed by mutation of the J-domain protein required for chloroplast accumulation response 1 (JAC1) gene, which is essential for the accumulation response; these results indicate that WEB1 and PMI2 play a role in suppressing JAC1 under strong light conditions. A yeast two-hybrid analysis and a nuclear recruitment assay identified a physical interaction between WEB1 and PMI2, and transient expression analysis of CFP-WEB1 and YFP-PMI2 revealed that they colocalized in the cytosol. Bimolecular fluorescence complementation analysis confirmed the interaction of these proteins in the cytosol. Blue light-induced changes in short chloroplast actin filaments (cp-actin filaments) were impaired in both web1 and pmi2. Our findings suggest that a cytosolic WEB1–PMI2 complex maintains the velocity of chloroplast photorelocation movement via cp-actin filament regulation.
机译:叶绿体向弱光(累积响应)移动,并远离强光(回避响应)。叶绿体在环境光条件下快速而准确的运动对于有效的光合作用和防止叶绿体的光损伤至关重要。在这里,我们报告说,两个拟南芥突变体,弱蓝绿素在蓝光1(web1)和web2下的运动在避免和积累响应方面均存在缺陷。基于图的克隆显示这两个基因都编码卷曲螺旋蛋白,并且WEB2与质体运动受损2(PMI2)基因相同。 web1和pmi2中叶绿体运动的速度大约比野生型低三倍。通过对叶绿体累积反应1(JAC1)基因所需的J结构域蛋白进行突变,可以抑制web1和pmi2回避反应的缺陷,这对于累积反应至关重要。这些结果表明,WEB1和PMI2在强光条件下起抑制JAC1的作用。酵母双杂交分析和核募集分析确定了WEB1和PMI2之间的物理相互作用,对CFP-WEB1和YFP-PMI2的瞬时表达分析表明它们共定位在胞质溶胶中。双分子荧光互补分析证实了这些蛋白在细胞质中的相互作用。蓝光诱导的短叶绿体肌动蛋白丝(cp-actin丝)的变化在web1和pmi2中均受到损害。我们的发现表明,胞质WEB1-PMI2复合物通过cp-actin细丝调节维持叶绿体光定位运动的速度。

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