Clustering and negative feedback by endocytosis in planar cell polarity signaling is modulated by ubiquitinylation of prickle.
Clustering and negative feedback by endocytosis in planar cell polarity signaling is modulated by ubiquitinylation of prickle.
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DOI：
10.1371/journal.pgen.1005259
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发表时间：
2015-05
期刊：
影响因子：
4.5
通讯作者：
Axelrod JD
中科院分区：
文献类型：
--
作者：
Cho B;Pierre-Louis G;Sagner A;Eaton S;Axelrod JD
平面细胞极性（PCP）信号系统的核心成分，包括跨膜和外周膜相关蛋白，形成桥接顶端细胞间连接的不对称复合物。虽然这些可以在任一方向上组装，但协调的细胞极化需要在特定连接处富集给定方向的复合物。这可能通过相反取向的复合物之间的正反馈和负反馈发生，并且需要外周膜相关的PCP组分。然而，反馈背后的分子机制还不清楚。我们发现，E3泛素连接酶复合物Cullin 1（Cul 1）/SkpA/多余肢（Slimb）调节外周膜成分之一，刺痛（Pk）的稳定性。过量的Pk破坏PCP反馈并防止不对称。我们发现，PK参与负反馈介导的PCP复合物的内化含有跨膜组件货车高（旺）和火烈鸟（FMI），和内化被激活的集群内的相反方向的复合物。Pk还通过一种未知的促进聚类的机制参与正反馈。因此，我们的研究结果确定了PCP信号不对称产生的分子机制。许多上皮细胞显示出一定水平的组织，其中细胞结构或附属物在细胞内沿着垂直于细胞的顶基轴的轴不对称地定位。当这种极化的方向在上皮平面内协调时，这种现象被称为平面细胞极性（PCP）。PCP至少部分由一组分子组成，这些分子在细胞-细胞连接处相互作用，并分离成两组，定位在每个细胞的相对两侧。它们的不对称定位被认为既产生分子不对称性，又标记细胞内的极化域，用于随后的形态极化。在分离以产生分子不对称性时，这些蛋白质参与正反馈和负反馈，就像铁磁体一样，以对齐它们在相邻细胞内和相邻细胞之间的定位。在这项工作中，我们确定了一种负反馈机制，利用蛋白质刺，PCP信号组件之一。Prickle的水平受到精确的调节，部分是通过泛素化机制来降解多余的蛋白质。刺痒感介导一类PCP蛋白质的内化和去除，从而引起相反“两极”的排斥。过量的刺痛会破坏这一机制，并干扰极性的建立。
The core components of the planar cell polarity (PCP) signaling system, including both transmembrane and peripheral membrane associated proteins, form asymmetric complexes that bridge apical intercellular junctions. While these can assemble in either orientation, coordinated cell polarization requires the enrichment of complexes of a given orientation at specific junctions. This might occur by both positive and negative feedback between oppositely oriented complexes, and requires the peripheral membrane associated PCP components. However, the molecular mechanisms underlying feedback are not understood. We find that the E3 ubiquitin ligase complex Cullin1(Cul1)/SkpA/Supernumerary limbs(Slimb) regulates the stability of one of the peripheral membrane components, Prickle (Pk). Excess Pk disrupts PCP feedback and prevents asymmetry. We show that Pk participates in negative feedback by mediating internalization of PCP complexes containing the transmembrane components Van Gogh (Vang) and Flamingo (Fmi), and that internalization is activated by oppositely oriented complexes within clusters. Pk also participates in positive feedback through an unknown mechanism promoting clustering. Our results therefore identify a molecular mechanism underlying generation of asymmetry in PCP signaling. Many epithelial cells display a level of organization in which cellular structures or appendages are positioned asymmetrically within the cell along an axis perpendicular to the apical-basal axis of the cell. When the direction of this polarization is coordinated within the plane of the epithelium, this phenomenon is referred to as planar cell polarity (PCP). PCP is organized, at least in part, by a group of molecules that interact across cell-cell junctions and segregate into two groups that localize on opposite sides of each cell. Their asymmetric localization is thought to both produce molecular asymmetry, and to mark polarized domains within the cell for subsequent morphological polarization. In segregating to produce molecular asymmetry, these proteins participate in both positive and negative feedback, much like ferromagnets, to align their localization within and between neighboring cells. In this work, we identify a mechanism for negative feedback that utilizes the protein Prickle, one of the PCP signaling components. Levels of Prickle are precisely regulated, in part by a ubiquitinylation mechanism that targets excess protein for degradation. Prickle mediates internalization and removal of one class of PCP proteins, thereby causing repulsion of opposite ‘poles.’ Excess Prickle disrupts this mechanism and interferes with establishing polarity.
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影响因子：
56.9
作者：
Amonlirdviman, K;Khare, NA;Tomlin, CJ
通讯作者：
Tomlin, CJ
影响因子：
11.8
作者：
Chin, Mei-Ling;Mlodzik, Marek
通讯作者：
Mlodzik, Marek
影响因子：
4.6
作者：
Das, G;Jenny, A;Mlodzik, M
通讯作者：
Mlodzik, M
影响因子：
11.8
作者：
Feiguin, F;Hannus, M;Eaton, S
通讯作者：
Eaton, S
DOI：
10.3791/1293
发表时间：
2009-08-19
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Choo, Yeun Su;Zhang, Zhuohua
通讯作者：
Zhang, Zhuohua