RhoA and membrane fluidity mediates the spatially polarized Src/FAK activation in response to shear stress.
RhoA and membrane fluidity mediates the spatially polarized Src/FAK activation in response to shear stress.
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RhoA 和膜流动性介导响应剪切应力的空间偏振 Src/FAK 激活
DOI：
10.1038/srep07008
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发表时间：
2014-11-12
影响因子：
4.6
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Liu B;Lu S;Hu YL;Liao X;Ouyang M;Wang Y
虽然Src在剪切应力诱导的细胞过程中起着至关重要的作用，但对高剪切应力（HSS）诱导的Src激活的时空模式知之甚少。HSS（65 dyn/cm 2）施加在牛主动脉内皮细胞上，利用基于荧光共振能量转移（FRET）的膜靶向Src生物传感器（Kras-Src）在亚细胞水平上观察动态Src激活。 极化Src激活观察到较高的活动在一侧面临的流动，这是增强了细胞松弛素D介导的破坏肌动蛋白丝，但抑制苯甲醇介导的膜流动性的增强。进一步的实验表明，HSS降低RhoA活性，与组成型活性RhoA突变体抑制，而负RhoA突变体增强HSS诱导的Src极性。细胞松弛素D可以恢复表达活性RhoA突变体的细胞的极性。进一步的结果表明，HSS刺激FAK激活的空间极性类似于Src。PP 1对Src的抑制以及RhoA活性和膜流动性的扰动可以阻断HSS诱导的FAK极性。这些结果表明，HSS诱导的Src和随后FAK极性依赖于由RhoA调节的细胞内张力分布，其相关的肌动蛋白结构和质膜流动性之间的协调。
While Src plays crucial roles in shear stress-induced cellular processes, little is known on the spatiotemporal pattern of high shear stress (HSS)-induced Src activation. HSS (65 dyn/cm2) was applied on bovine aortic endothelial cells to visualize the dynamic Src activation at subcellular levels utilizing a membrane-targeted Src biosensor (Kras-Src) based on fluorescence resonance energy transfer (FRET). A polarized Src activation was observed with higher activity at the side facing the flow, which was enhanced by a cytochalasin D-mediated disruption of actin filaments but inhibited by a benzyl alcohol-mediated enhancement of membrane fluidity. Further experiments revealed that HSS decreased RhoA activity, with a constitutively active RhoA mutant inhibiting while a negative RhoA mutant enhancing the HSS-induced Src polarity. Cytochalasin D can restore the polarity in cells expressing the active RhoA mutant. Further results indicate that HSS stimulates FAK activation with a spatial polarity similar to Src. The inhibition of Src by PP1, as well as the perturbation of RhoA activity and membrane fluidity, can block this HSS-induced FAK polarity. These results indicate that the HSS-induced Src and subsequently FAK polarity depends on the coordination between intracellular tension distribution regulated by RhoA, its related actin structures and the plasma membrane fluidity.
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影响因子：
21.3
作者：
Felsenfeld, DP;Schwartzberg, PL;Sheetz, MP
通讯作者：
Sheetz, MP
DOI：
10.1083/jcb.153.7.1427
发表时间：
2001-06-25
期刊：
The Journal of cell biology
影响因子：
--
作者：
Laukaitis CM;Webb DJ;Donais K;Horwitz AF
通讯作者：
Horwitz AF
DOI：
10.1152/ajpheart.01156.2007
发表时间：
2008-08-01
影响因子：
4.8
作者：
Metaxa, Eleni;Meng, Hui;Kolega, John
通讯作者：
Kolega, John
影响因子：
15.9
作者：
Li, S;Chen, BPC;Chien, S
通讯作者：
Chien, S
影响因子：
4.8
作者：
Lunn, JA;Rozengurt, E
通讯作者：
Rozengurt, E