Proximity labeling reveals non-centrosomal microtubule-organizing center components required for microtubule growth and localization.
Proximity labeling reveals non-centrosomal microtubule-organizing center components required for microtubule growth and localization.
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DOI：
10.1016/j.cub.2021.06.021
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发表时间：
2021-08-23
期刊：
影响因子：
--
通讯作者：
Feldman JL
中科院分区：
文献类型：
--
作者：
Sanchez AD;Branon TC;Cote LE;Papagiannakis A;Liang X;Pickett MA;Shen K;Jacobs-Wagner C;Ting AY;Feldman JL
微管是一种极化的细胞内聚合物，在细胞的运输、极性和细胞分裂中发挥关键作用。在真核细胞类型中，微管采用不同的细胞内组织来适应这些不同的功能，这些不同的功能由称为微管组织中心(MTOCs)的特定细胞位置协调。50多年来，对MTOC生物学的研究主要集中在中心体上，然而大多数分化的细胞利用非中心体MTOCs(NcMTOCs)将其微管组织成不同的阵列，这对细胞功能至关重要。为了确定ncMTOC的基本成分，我们开发了基于生物素连接酶的邻近标记方法TurboID用于线虫。我们在肠上皮细胞顶端ncMTOC处发现了微管近端减去末端蛋白PTRN-1/Patronin的蛋白，重点分析了两个保守的蛋白：spectraplakin蛋白VAB-10B和WDR-62，我们发现该蛋白与脊椎动物原发小头症蛋白WDR62同源。VAB-10B和WDR-62不与中心体相关，而是专门调节非中心体微管和微管末端蛋白的顶端靶向。VAB-10B的缺失导致微管定位错误和微管成核复合体γ-TURC的定位延迟，而WDR-62的缺失导致动态微管数量减少和γ-TURC定位取消。这种调节发生在细胞极性的下游，并与肌动蛋白结合。由于这是首次报道WDR62家族蛋白的非中心体作用，我们扩展了这一重要疾病蛋白的基本细胞生物学作用。我们的研究确定了ncMTOC的基本成分，并提出了微管生长和定位受到明显调控的劳动分工。分化细胞利用非中心体微管组织中心(NcMTOCs)构建微管阵列。Sanchez等人。将邻近标记应用于活的线虫以生成ncMTOC的第一个蛋白质组图谱。NcMTOC组分的耗尽揭示了控制微管生长和定位的不同功能模块。
Microtubules are polarized intracellular polymers that play key roles in the cell including in transport, polarity, and cell division. Across eukaryotic cell types, microtubules adopt diverse intracellular organization to accommodate these distinct functions coordinated by specific cellular sites called microtubule-organizing centers (MTOCs). Over 50 years of research on MTOC biology has focused mainly on the centrosome, however most differentiated cells employ non-centrosomal MTOCs (ncMTOCs) to organize their microtubules into diverse arrays which are critical to cell function. To identify essential ncMTOC components, we developed the biotin ligase-based proximity labeling approach TurboID for use in C. elegans. We identified proteins proximal to the microtubule minus end protein PTRN-1/Patronin at the apical ncMTOC of intestinal epithelial cells, focusing on two conserved proteins: spectraplakin protein VAB-10B and WDR-62, a protein we identify as homologous to vertebrate primary microcephaly disease protein WDR62. VAB-10B and WDR-62 do not associate with the centrosome and instead specifically regulate non-centrosomal microtubules and the apical targeting of microtubule minus end proteins. Depletion of VAB-10B resulted in microtubule mislocalization and delayed localization of a microtubule nucleation complex γ-TuRC, while loss of WDR-62 decreased the number of dynamic microtubules and abolished γ-TuRC localization. This regulation occurs downstream of cell polarity and in conjunction with actin. As this is the first report for non-centrosomal roles of WDR62 family proteins, we expand the basic cell biological roles of this important disease protein. Our studies identify essential ncMTOC components and suggest a division of labor where microtubule growth and localization are distinctly regulated. Differentiated cells use non-centrosomal microtubule organizing centers (ncMTOCs) to build microtubule arrays. Sanchez et al. apply proximity labeling in living C. elegans to generate the first proteomic profile of an ncMTOC. Depletion of ncMTOC components reveals functionally distinct modules controlling microtubule growth and localization.
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影响因子：
3.3
作者：
Applewhite DA;Grode KD;Keller D;Zadeh AD;Slep KC;Rogers SL
通讯作者：
Rogers SL
影响因子：
64.8
作者：
Harris CR;Millman KJ;van der Walt SJ;Gommers R;Virtanen P;Cournapeau D;Wieser E;Taylor J;Berg S;Smith NJ;Kern R;Picus M;Hoyer S;van Kerkwijk MH;Brett M;Haldane A;Del Río JF;Wiebe M;Peterson P;Gérard-Marchant P;Sheppard K;Reddy T;Weckesser W;Abbasi H;Gohlke C;Oliphant TE
通讯作者：
Oliphant TE
DOI：
10.1083/jcb.200202047
发表时间：
2002-05-13
期刊：
The Journal of cell biology
影响因子：
--
作者：
Hannak E;Oegema K;Kirkham M;Gönczy P;Habermann B;Hyman AA
通讯作者：
Hyman AA
影响因子：
10.7
作者：
Baines, Anthony J.;Bignone, Paola A.;Phillips, Gareth W.
通讯作者：
Phillips, Gareth W.
影响因子：
4
作者：
Gupta, Kamlesh K.;Alberico, Emily O.;Goodson, Holly V.
通讯作者：
Goodson, Holly V.