Hypoxia‐stimulated membrane trafficking requires T‐plastin
Hypoxia‐stimulated membrane trafficking requires T‐plastin
复制标题
缺氧刺激的膜运输需要 Tâplastin
DOI：
10.1111/apha.12859
复制
发表时间：
2017
影响因子：
6.3
通讯作者：
Böttger J.
中科院分区：
文献类型：
--
作者：
Wottawa M;Naas S;Böttger J.
真核细胞的质膜和内膜间室之间的交通是真核细胞的基本特征。分泌途径将货物从生物合成室运送到质膜。这是由逆行的内吞途径平衡的，是细胞平衡所必需的。细胞需要快速适应环境挑战，如po2的减少，然而，尚未详细分析与膜运输有关的环境挑战。因此，我们确定了常氧、缺氧和再氧合后质膜运输的变化。方法采用大体积膜内吞标志物fm1‐43、新开发的膜探针mCLING、小麦胚芽凝集素以及荧光标记的霍乱毒素亚基b来分析膜运输，并测定特异性膜蛋白的摄取。同时，进行无偏差SILAC筛选，分析在常氧和缺氧条件下膜蛋白的丰度。结果缺氧时膜转运增加，再氧后迅速逆转。这种效应与缺氧诱导因子（HIF）系统无关。利用SILAC技术，我们发现肌动蛋白捆绑蛋白T - plastin在缺氧条件下被招募到质膜上。通过使用T - plastin敲除细胞，我们可以证明T - plastin介导缺氧诱导的膜运输，这与细胞中肌动蛋白密度的增加有关。结论缺氧时膜转运具有高度的动态性。这种表型在再氧化后迅速可逆，这表明这种机制参与了细胞对缺氧的适应。
AimTraffic between the plasma membrane and the endomembrane compartments is an essential feature of eukaryotic cells. The secretory pathway sends cargoes from biosynthetic compartments to the plasma membrane. This is counterbalanced by a retrograde endocytic route and is essential for cell homoeostasis. Cells need to adapt rapidly to environmental challenges such as the reduction of pO2which, however, has not been analysed in relation to membrane trafficking in detail. Therefore, we determined changes in the plasma membrane trafficking in normoxia, hypoxia, and after reoxygenation.MethodsMembrane trafficking was analysed by using the bulk membrane endocytosis marker FM 1‐43, the newly developed membrane probe mCLING, wheat germ agglutinin as well as fluorescently labelled cholera toxin subunit B. Additionally, the uptake of specific membrane proteins was determined. In parallel, a non‐biased SILAC screen was performed to analyse the abundance of membrane proteins in normoxia and hypoxia.ResultsMembrane trafficking was increased in hypoxia and quickly reversed upon reoxygenation. This effect was independent of the hypoxia‐inducible factor (HIF) system. Using SILAC technology, we identified that the actin‐bundling protein T‐plastin is recruited to the plasma membrane in hypoxia. By the use of T‐plastin knockdown cells, we could show that T‐plastin mediates the hypoxia‐induced membrane trafficking, which was associated with an increased actin density in the cells as determined by electron microscopy.ConclusionMembrane trafficking is highly dynamic upon hypoxia. This phenotype is quickly reversible upon reoxygenation, which suggests that this mechanism participates in the cellular adaptation to hypoxia.
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整体制备和切片中的肌动蛋白细胞骨架。
影响因子：
--
作者：
G. Resch;E. Urbán;S. Jacob
通讯作者：
S. Jacob
影响因子：
32.4
作者：
Chen, H;Mocsai, A;Brown, EJ
通讯作者：
Brown, EJ
影响因子：
5.3
作者：
Wang Y;Ohh M
通讯作者：
Ohh M
DOI：
10.1083/jcb.201402066
发表时间：
2014-05-26
期刊：
The Journal of cell biology
影响因子：
--
作者：
Revelo NH;Kamin D;Truckenbrodt S;Wong AB;Reuter-Jessen K;Reisinger E;Moser T;Rizzoli SO
通讯作者：
Rizzoli SO
内吞途径的电子显微镜。
影响因子：
--
作者：
C. Ranftler;Peter Auinger;C. Meisslitzer;A. Ellinger;J. Neumüller;M. Pavelka
通讯作者：
M. Pavelka