Dynamic relationship of focal contacts and hemidesmosome protein complexes in live cells.
Dynamic relationship of focal contacts and hemidesmosome protein complexes in live cells.
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DOI：
10.1038/jid.2009.439
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发表时间：
2010-06
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影响因子：
--
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中科院分区：
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--
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表皮细胞通过称为半丝粒的细胞-基质连接粘附在基底膜区。在伤口愈合过程中，半底粒被分解，使角化细胞能够在伤口上移动。这种运动是由半粒酶体蛋白复合物（HPCs）和局灶接触（fc）介导的。本研究在以黄色荧光蛋白（YFP）标记的β4整合素和青色荧光蛋白（CFP）标记的α-肌动蛋白分别作为HPCs和FCs标记的活HaCat细胞中，分析了HPCs和FCs之间的相互作用。在HaCat细胞迁移到伤口的过程中，FC蛋白在伤口的方向上迅速聚集。然后是HPC组装，新形成的HPC占据被拆卸的fc腾出的位置。HPC动力学显著降低，HaCat细胞在使用影响FC完整性/功能的试剂处理后停止迁移。在用破坏HPCs稳定性的试剂处理后，伤口边缘HaCat细胞中FCs的动力学得到增强，尽管FC的组装是不规则的，细胞的迁移是异常的。我们还表明，角化细胞中半脂粒和fc之间的复杂相互作用依赖于肌球蛋白，需要能量。总之，我们认为在伤口愈合过程中，HPCs和FCs的动态在角化细胞迁移过程中受到密切的共同调节。
Epidermal cells adhere to the basement membrane zone through cell–matrix junctions termed hemidesmosomes. During wound healing, hemidesmosomes are disassembled to allow keratinocytes to move over wound sites. Such movement is mediated by both hemidesmosome protein complexes (HPCs) and focal contacts (FCs). In this study, we analyzed the interaction between HPCs and FCs in live HaCat cells expressing yellow fluorescent protein (YFP)-tagged β4 integrin and cyan fluorescent protein (CFP)-tagged α-actinin as markers of HPCs and FCs, respectively. In HaCat cells migrating to repopulate wounds, FC proteins cluster rapidly in the direction of the wound. HPC assembly then follows and the newly formed HPCs occupy sites vacated by the disassembled FCs. HPC dynamics are dramatically reduced, and HaCat cells cease migration upon treatment with reagents that affect FC integrity/function. Upon treatment with reagents that destabilize HPCs, the dynamics of FCs in HaCat cells at the edges of wounds are enhanced, although FC assembly is irregular and the migration of the cells is aberrant. We also show that the complex interaction between hemidesmosomes and FCs in keratinocytes is myosin dependent and requires energy. In summary, we suggest that HPCs and FCs dynamics are tightly co-regulated in keratinocytes undergoing migration during wound healing.
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