Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells.
Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells.
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DOI：
10.1083/jcb.153.7.1427
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发表时间：
2001-06-25
期刊：
影响因子：
--
通讯作者：
Horwitz AF
中科院分区：
文献类型：
--
作者：
Laukaitis CM;Webb DJ;Donais K;Horwitz AF
为了研究粘附在迁移细胞中形成和分散的机制，我们表达了α5整合素、α-辅肌动蛋白和桩蛋白作为绿色荧光蛋白（GFP）融合体。所有本地化的内源性对应物，并没有干扰迁移时，在中等水平表达。α5-GFP还挽救了CHO B2细胞中的粘附缺陷，该细胞是α5整联蛋白缺陷的。在皱褶细胞中，α5-GFP和α-actinin-GFP显著定位于膜突起的前缘。在我们检查的三种GFP融合蛋白中，桩蛋白是第一个在细胞突起区域中出现明显组织的成分。当形成新的突出物时，桩蛋白似乎在前缘从旧的粘连重塑为新的粘连。α-辅肌动蛋白随后进入粘连，其向细胞中心移位，并抑制桩蛋白更新。新的粘连由α-辅肌动蛋白-GFP和paxillin-GFP的小病灶形成，其尺寸增大。随后，α5整联蛋白进入粘连形成可见的复合物，起到稳定粘连的作用。α5-GFP也存在于从突起前缘发出的内吞囊泡中。细胞后部的整合素囊泡向细胞体移动。随着细胞的迁移，α5囊泡也从核周区域移动到板状伪足的基部。α5囊泡与转铁蛋白受体和FM 4-64染料共定位。在后部粘连破裂后，在细胞后面的纤维结构中发现α5-GFP，而α-actinin-GFP和paxillin-GFP作为有组织的结构向上移动到缩回细胞的侧边缘，然后消散。
To investigate the mechanisms by which adhesions form and disperse in migrating cells, we expressed α5 integrin, α-actinin, and paxillin as green fluorescent protein (GFP) fusions. All localized with their endogenous counterparts and did not perturb migration when expressed at moderate levels. α5-GFP also rescued the adhesive defects in CHO B2 cells, which are α5 integrin deficient. In ruffling cells, α5-GFP and α-actinin–GFP localized prominently at the leading edge in membrane protrusions. Of the three GFP fusion proteins that we examined, paxillin was the first component to appear visibly organized in protrusive regions of the cell. When a new protrusion formed, the paxillin appeared to remodel from older to newer adhesions at the leading edge. α-Actinin subsequently entered adhesions, which translocated toward the cell center, and inhibited paxillin turnover. The new adhesions formed from small foci of α-actinin–GFP and paxillin-GFP, which grew in size. Subsequently, α5 integrin entered the adhesions to form visible complexes, which served to stabilize the adhesions. α5-GFP also resided in endocytic vesicles that emanated from the leading edge of protrusions. Integrin vesicles at the cell rear moved toward the cell body. As cells migrated, α5 vesicles also moved from a perinuclear region to the base of the lamellipodium. The α5 vesicles colocalized with transferrin receptor and FM 4-64 dye. After adhesions broke down in the rear, α5-GFP was found in fibrous structures behind the cell, whereas α-actinin–GFP and paxillin-GFP moved up the lateral edge of retracting cells as organized structures and then dissipated.
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影响因子：
7.8
作者：
Borowsky, ML;Hynes, RO
通讯作者：
Hynes, RO
影响因子：
56.9
作者：
MIYAMOTO, S;AKIYAMA, SK;YAMADA, KM
通讯作者：
YAMADA, KM
影响因子：
7.8
作者：
DePasquale, J A;Izzard, C S
通讯作者：
Izzard, C S
DOI：
10.1016/0922-3371(90)90048-2
发表时间：
1990-12-01
期刊：
CELL DIFFERENTIATION AND DEVELOPMENT
影响因子：
--
作者：
BURRIDGE, K;NUCKOLLS, G;TURNER, C
通讯作者：
TURNER, C
DOI：
10.1073/pnas.94.17.9493
发表时间：
1997-08-19
影响因子：
11.1
作者：
Dabiri, GA;Turnacioglu, KK;Sanger, JW
通讯作者：
Sanger, JW