Clustering of desmosomal cadherins by desmoplakin is essential for cell-cell adhesion
Clustering of desmosomal cadherins by desmoplakin is essential for cell-cell adhesion
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桥粒斑蛋白对桥粒钙粘蛋白的聚集对于细胞与细胞的粘附至关重要
DOI：
10.1111/apha.13609
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Spindler V
中科院分区：
文献类型：
--
作者：
Wanuske MT;Brantschen D;Schinner C;Stüdle C;Walter E;Hiermaier M;Vielmuth F;Waschke J;Spindler V
桥粒蛋白(DP)是桥粒的重要组成部分，桥粒是锚定中间丝的超分子细胞连接复合体。DP如何调节细胞间黏附的机制还只有部分了解。在此，我们研究了DP对桥粒黏附分子功能、桥粒转换率和细胞间黏附的影响。方法用免疫印迹和免疫染色方法对人角质形成细胞进行基因编辑。用电子显微镜和细胞黏附实验检测桥粒的超微结构和功能。用原子力显微镜和超分辨成像研究了桥粒钙粘附素的单分子结合特性和定位。结果DP的KO敲除(KO)与另一种桥粒蛋白Pg的KO一样，显著降低了细胞的凝聚力。与PG Ko相反，DP Ko完全缺乏桥粒。桥粒黏附分子Desmocolin(DSC)3和Desmoglein(DSG)3的结合特性在DP丢失时保持不变。由于Dsg2和Dsc3主要定位于桥粒内的黏附分子Dsg2和Dsc3被重新分布到DP Ko细胞膜上的小斑点，因此DP是将桥粒钙粘附素组装成大簇所必需的。DP KO中桥粒钙粘附素的额外沉默不会进一步增加细胞间黏附的丧失。结论：数据表明DP对桥粒的形成是必要的，但不影响单个钙粘附素结合特性水平上的细胞间黏附。相反，通过DP对桥粒黏附分子进行宏观聚集是至关重要的。这些结果可能有助于更好地了解由DP功能障碍引起的严重疾病。
AIMDesmoplakin (Dp) is a crucial component of the desmosome, a supramolecular cell junction complex anchoring intermediate filaments. The mechanisms how Dp modulates cell‐cell adhesion are only partially understood. Here, we studied the impact of Dp on the function of desmosomal adhesion molecules, desmosome turnover and intercellular adhesion.METHODSCRISPR/Cas9 was used for gene editing of human keratinocytes which were characterized by Western blot and immunostaining. Desmosomal ultrastructure and function were assessed by electron microscopy and cell adhesion assays. Single molecule binding properties and localization of desmosomal cadherins were studied by atomic force microscopy and super‐resolution imaging.RESULTSKnockout (ko) of Dp impaired cell cohesion to drastically higher extents as ko of another desmosomal protein, plakoglobin (Pg). In contrast to Pg ko, desmosomes were completely absent in Dp ko. Binding properties of the desmosomal adhesion molecules desmocollin (Dsc) 3 and desmoglein (Dsg) 3 remained unaltered under loss of Dp. Dp was required for assembling desmosomal cadherins into large clusters, as Dsg2 and Dsc3, adhesion molecules primarily localized within desmosomes, were redistributed into small puncta in the cell membrane of Dp ko cells. Additional silencing of desmosomal cadherins in Dp ko did not further increase loss of intercellular adhesion.CONCLUSIONOur data demonstrate that Dp is essential for desmosome formation but does not influence intercellular adhesion on the level of individual cadherin binding properties. Rather, macro‐clustering of desmosomal adhesion molecules through Dp is crucial. These results may help to better understand severe diseases which are caused by Dp dysfunction.
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影响因子：
4.6
作者：
Huetz, Katharina;Zeiler, Julian;Spindler, Volker
通讯作者：
Spindler, Volker
桥粒斑块的主要多肽桥粒斑蛋白 I 和 II 的生化和免疫学特征。
影响因子：
5.6
作者：
Helga Mueller;W. W. Franke
通讯作者：
W. W. Franke
角蛋白通过信号传导调节桥粒钙粘蛋白的粘附特性
DOI：
10.1016/j.jid.2017.08.033
发表时间：
2018
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
Vielmuth F;Wanuske MT;Radeva MY;Hiermaier M;Kugelmann D;Walter E;Buechau F;Magin TM;Waschke J;Spindler V
通讯作者：
Spindler V
影响因子：
4
作者：
Cabral, Rita M.;Tattersall, Daniel;Kelsell, David P.
通讯作者：
Kelsell, David P.
DOI：
10.1038/jid.2014.21
发表时间：
2014-06
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
V. Spindler;C. Dehner;S. Hübner;J. Waschke
通讯作者：
V. Spindler;C. Dehner;S. Hübner;J. Waschke