Optogenetics-based localization of talin to the plasma membrane promotes activation of β3 integrins.
Optogenetics-based localization of talin to the plasma membrane promotes activation of β3 integrins.
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DOI：
10.1016/j.jbc.2021.100675
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Shattil SJ
中科院分区：
文献类型：
--
作者：
Liao Z;Gingras AR;Lagarrigue F;Ginsberg MH;Shattil SJ
talin与整联蛋白β的胞质尾的相互作用触发整联蛋白活化，导致整联蛋白对细胞外配体的亲和力/亲合力增加。在talin KO小鼠中，talin与血小板整合素αIIbβ3相互作用的丧失导致严重的止血缺陷，而talin与内皮细胞整合素αVβ3相互作用的丧失影响血管生成。在正常细胞中，talin被自身抑制并定位于细胞质中。在这里，我们使用光遗传学平台来评估全长talin到质膜的募集是否足以诱导整合素活化。将对450 nm（蓝色）光响应的二聚化模块（拟南芥隐花色素2融合到talin的N末端;隐花色素相互作用的碱性螺旋-环-螺旋结构域的N末端以CAAX盒蛋白[C：半胱氨酸; A：脂肪族氨基酸; X：任何C末端氨基酸]结束）分别插入到也表达αIIbβ3或αVβ3的中国仓鼠卵巢细胞和内皮细胞中。因此，细胞暴露于蓝光引起拟南芥隐花色素2-talin的快速和可逆的招聘到隐花色素相互作用的基本螺旋-环-螺旋结构域的N-末端，该结构域以CAAX盒蛋白[C：半胱氨酸; A：脂肪族氨基酸; X：任何C-末端氨基酸]修饰的质膜结束。这导致两种细胞类型中的β3整联蛋白活化，以及内皮细胞迁移增加。然而，膜募集塔林是不足以激活整合素，膜相关Ras相关蛋白1（Rap 1）-GTP也是必需的。此外，干扰其与Rap 1直接结合的talin突变废除了β3整合素活化。总之，这些结果确定了talin在β3整合素活化中的质膜募集作用，并且它们表明此后涉及Rap 1-GTP的事件的细微差别。
Interaction of talin with the cytoplasmic tails of integrin β triggers integrin activation, leading to an increase of integrin affinity/avidity for extracellular ligands. In talin KO mice, loss of talin interaction with platelet integrin αIIbβ3 causes a severe hemostatic defect, and loss of talin interaction with endothelial cell integrin αVβ3 affects angiogenesis. In normal cells, talin is autoinhibited and localized in the cytoplasm. Here, we used an optogenetic platform to assess whether recruitment of full-length talin to the plasma membrane was sufficient to induce integrin activation. A dimerization module (Arabidopsis cryptochrome 2 fused to the N terminus of talin; N-terminal of cryptochrome-interacting basic helix-loop-helix domain ended with a CAAX box protein [C: cysteine; A: aliphatic amino acid; X: any C-terminal amino acid]) responsive to 450 nm (blue) light was inserted into Chinese hamster ovary cells and endothelial cells also expressing αIIbβ3 or αVβ3, respectively. Thus, exposure of the cells to blue light caused a rapid and reversible recruitment of Arabidopsis cryptochrome 2–talin to the N-terminal of cryptochrome-interacting basic helix-loop-helix domain ended with a CAAX box protein [C: cysteine; A: aliphatic amino acid; X: any C-terminal amino acid]–decorated plasma membrane. This resulted in β3 integrin activation in both cell types, as well as increasing migration of the endothelial cells. However, membrane recruitment of talin was not sufficient for integrin activation, as membrane-associated Ras-related protein 1 (Rap1)–GTP was also required. Moreover, talin mutations that interfered with its direct binding to Rap1 abrogated β3 integrin activation. Altogether, these results define a role for the plasma membrane recruitment of talin in β3 integrin activation, and they suggest a nuanced sequence of events thereafter involving Rap1–GTP.
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影响因子：
48
作者：
Kennedy, Matthew J.;Hughes, Robert M.;Peteya, Leslie A.;Schwartz, Joel W.;Ehlers, Michael D.;Tucker, Chandra L.
通讯作者：
Tucker, Chandra L.
DOI：
10.1038/nrm3255
发表时间：
2011-12-22
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1073/pnas.87.22.8960
发表时间：
1990-11-01
影响因子：
11.1
作者：
KAWATA, M;FARNSWORTH, CC;TAKAI, Y
通讯作者：
TAKAI, Y
影响因子：
7.8
作者：
Beckerle, M C;Miller, D E;Bertagnolli, M E;Locke, S J
通讯作者：
Locke, S J
影响因子：
3.7
作者：
Kahner BN;Kato H;Banno A;Ginsberg MH;Shattil SJ;Ye F
通讯作者：
Ye F