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首页> 外文期刊>Molecular Microbiology >Apical assemblies of intermediate filament‐like protein FilP are highly dynamic and affect polar growth determinant DivIVA in Streptomyces venezuelae Streptomyces venezuelae
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Apical assemblies of intermediate filament‐like protein FilP are highly dynamic and affect polar growth determinant DivIVA in Streptomyces venezuelae Streptomyces venezuelae

机译:中间丝状蛋白质Filp的顶端组件是高度动态的,并且影响乙烯脲链霉菌委内瑞斯乙烯脲酶中的极性生长决定因子Diviva

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摘要

Summary Streptomyces spp. grow as branching hyphae, building the cell wall in restricted zones at hyphal tips. The organization of this mode of polar growth involves three coiled‐coil proteins: DivIVA and Scy, which form apical protein complexes referred to as polarisomes; and the intermediate filament‐like protein FilP, which influences cell shape and interacts with both Scy and DivIVA. Here, we use live cell imaging of Streptomyces venezuelae to clarify the subcellular localization and dynamics of FilP and its effect on hyphal morphology. By monitoring a FilP‐mCherry fusion protein, we show that FilP accumulates in gradient‐like zones behind the hyphal tips. The apical gradient pattern of FilP localization is dependent on hyphal tip extension and immediately dissipates upon growth arrest. Fluorescence recovery after photobleaching experiments show that FilP gradients are dynamic and subject to subunit exchange during vegetative growth. Further, the localization of FilP at hyphal tips is not directly dependent on scy , even though the strongly perturbed morphology of most scy mutant hyphae is associated with mislocalization of FilP. Finally, we find that filP has an effect on the size and position of the foci of key polar growth determinant DivIVA. This effect likely contributes to the phenotype of filP mutants.
机译:摘要Streptomyces SPP。成长为分枝菌丝,在悬垂尖端的限制区建造细胞壁。这种极地生长模式的组织涉及三种卷曲卷蛋白:Diviva和Scy,其形成称为极热体的顶端蛋白质复合物;和中间丝状蛋白质Filp,其影响细胞形状并与Scy和Diviva相互作用。在这里,我们使用Streptomyces委内瑞拉的活细胞成像,阐明Filp的亚细胞定位和动态及其对亚酚形态的影响。通过监测Filp-MCHerry融合蛋白,我们显示FILP累积在剪码提示后面的梯度状区域。 FILP定位的顶端梯度模式取决于悬垂尖端延伸,并立即在生长停止时耗散。光漂白实验后的荧光回收表明,在营养生长期间,Filp梯度是动态的,受亚基交换。此外,即使大多数Scy突变体菌株的强烈扰动形态与Filp的错误分析相关,悬角提示的悬角尖端局部的定位也不直接依赖于SCY。最后,我们发现Filp对关键极性生长决定因子Diviva的焦点的大小和位置有影响。这种效果可能有助于菲尔普突变体的表型。

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