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多价卷曲螺旋相互作用使中心体完全组装和增强。

Multivalent coiled-coil interactions enable full-scale centrosome assembly and strength.

机构信息

Department of Cell Biology, Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX, USA.

Department of Biochemistry, Center for Alzheimer's and Neurodegenerative Diseases, Peter O'Donnell Jr. Brain Institute, University of Texas Southwestern Medical Center, Dallas, TX, USA.

出版信息

J Cell Biol. 2024 Apr 1;223(4). doi: 10.1083/jcb.202306142. Epub 2024 Mar 8.

DOI:10.1083/jcb.202306142
PMID:38456967
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10921949/
Abstract

The outermost layer of centrosomes, called pericentriolar material (PCM), organizes microtubules for mitotic spindle assembly. The molecular interactions that enable PCM to assemble and resist external forces are poorly understood. Here, we use crosslinking mass spectrometry (XL-MS) to analyze PLK-1-potentiated multimerization of SPD-5, the main PCM scaffold protein in C. elegans. In the unassembled state, SPD-5 exhibits numerous intramolecular crosslinks that are eliminated after phosphorylation by PLK-1. Thus, phosphorylation induces a structural opening of SPD-5 that primes it for assembly. Multimerization of SPD-5 is driven by interactions between multiple dispersed coiled-coil domains. Structural analyses of a phosphorylated region (PReM) in SPD-5 revealed a helical hairpin that dimerizes to form a tetrameric coiled-coil. Mutations within this structure and other interacting regions cause PCM assembly defects that are partly rescued by eliminating microtubule-mediated forces, revealing that PCM assembly and strength are interdependent. We propose that PCM size and strength emerge from specific, multivalent coiled-coil interactions between SPD-5 proteins.

摘要

中心体的最外层称为中心粒周围物质（PCM），它组织有丝分裂纺锤体的微管组装。PCM 组装和抵抗外力的分子相互作用还知之甚少。在这里，我们使用交联质谱（XL-MS）分析 PLK-1 增强的 SPD-5 多聚化，SPD-5 是秀丽隐杆线虫中主要的 PCM 支架蛋白。在未组装状态下，SPD-5 表现出许多分子内交联，这些交联在被 PLK-1 磷酸化后被消除。因此，磷酸化诱导 SPD-5 的结构开放，使其为组装做好准备。SPD-5 的多聚化是由多个分散的卷曲螺旋结构域之间的相互作用驱动的。对 SPD-5 中一个磷酸化区域（PReM）的结构分析揭示了一个螺旋发夹，它二聚形成四聚卷曲螺旋。该结构内的突变和其他相互作用区域导致 PCM 组装缺陷，部分缺陷可以通过消除微管介导的力来挽救，这表明 PCM 组装和强度是相互依存的。我们提出，PCM 的大小和强度源自 SPD-5 蛋白之间特定的、多价的卷曲螺旋相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/51ba8cd146dd/JCB_202306142_FigS5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/7242997268b4/JCB_202306142_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/0c0174b7f497/JCB_202306142_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/48611218539f/JCB_202306142_FigS2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/fae2641e8dc3/JCB_202306142_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/b54463ca2e5e/JCB_202306142_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/274dc09b64dc/JCB_202306142_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/85d8449f1cda/JCB_202306142_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/51ba8cd146dd/JCB_202306142_FigS5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/7242997268b4/JCB_202306142_Fig1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/0c0174b7f497/JCB_202306142_FigS1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/48611218539f/JCB_202306142_FigS2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/fae2641e8dc3/JCB_202306142_Fig2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/b54463ca2e5e/JCB_202306142_Fig3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/274dc09b64dc/JCB_202306142_FigS3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/85d8449f1cda/JCB_202306142_Fig4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/375f/10921949/51ba8cd146dd/JCB_202306142_FigS5.jpg

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