The Intracranial Aneurysm Gene THSD1 Connects Endosome Dynamics to Nascent Focal Adhesion Assembly
| Autor: | Yanning Rui, John P. Hagan, Dong H. Kim, Xiaoqian Fang, Airu Niu, Julien Balzeau, Miriam R. Menezes, Zhen Xu |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Talin
0301 basic medicine Physiology Endosome Integrin Focal adhesion assembly Endosomes Bioinformatics lcsh:Physiology Zyxin Focal adhesion lcsh:Biochemistry 03 medical and health sciences Human Umbilical Vein Endothelial Cells Humans Immunoprecipitation lcsh:QD415-436 RNA Small Interfering Focal Adhesions biology lcsh:QP1-981 Chemistry Protein complex Intracranial Aneurysm Adhesion Vinculin Talin binding Clathrin Endocytosis Cell biology HEK293 Cells 030104 developmental biology Microscopy Fluorescence Focal Adhesion Kinase 1 biology.protein RNA Interference Thrombospondins Thsd1 HeLa Cells |
| Zdroj: | Cellular Physiology and Biochemistry, Vol 43, Iss 6, Pp 2200-2211 (2017) |
| ISSN: | 1421-9778 1015-8987 |
| Popis: | Background/Aims: We recently discovered that harmful variants in THSD1 (Thrombospondin type-1 domain-containing protein 1) likely cause intracranial aneurysm and subarachnoid hemorrhage in a subset of both familial and sporadic patients with supporting evidence from two vertebrate models. The current study seeks to elucidate how THSD1 and patient-identified variants function molecularly in focal adhesions. Methods: Co-immunostaining and co-immunoprecipitation were performed to define THSD1 subcellular localization and interacting partners. Transient expression of patient-identified THSD1 protein variants and siRNA-mediated loss-of-function THSD1 were used to interrogate gene function in focal adhesion and cell attachment to collagen I in comparison to controls. Results: THSD1 is a novel nascent adhesion protein that co-localizes with several known markers such as FAK, talin, and vinculin, but not with mature adhesion marker zyxin. Furthermore, THSD1 forms a multimeric protein complex with FAK/talin/vinculin, wherein THSD1 promotes talin binding to FAK but not to vinculin, a key step in nascent adhesion assembly. Accordingly, THSD1 promotes mature adhesion formation and cell attachment, while its rare variants identified in aneurysm patients show compromised ability. Interestingly, THSD1 also localizes at different stages of endosomes. Clathrin-mediated but not caveolae-mediated endocytosis pathway is involved in THSD1 intracellular trafficking, which positively regulates THSD1-induced focal adhesion assembly, in contrast to the traditional role of endosomes in termination of integrin signals. Conclusions: The data suggest that THSD1 functions at the interface between endosome dynamics and nascent focal adhesion assembly that is impaired by THSD1 rare variants identified from intracranial aneurysm patients. |
| Databáze: | OpenAIRE |
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