Live-cell imaging of the association of STAT6-GFP with mitochondria.
Live-cell imaging of the association of STAT6-GFP with mitochondria.
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DOI：
10.1371/journal.pone.0055426
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Sehgal PB
中科院分区：
文献类型：
--
作者：
Khan R;Lee JE;Yang YM;Liang FX;Sehgal PB
转录因子STAT 3先前已被报道与线粒体相关。然而，迄今为止，我们还不能观察到STAT 3-GFP、STAT 3-DsRed或STAT 3-Flag与人Hep 3B肝细胞中线粒体的关联，即使这些分子与其他细胞质细胞器（内体）的关联是容易证明的。然后，我们解决了一个更广泛的问题，其他STAT家族的蛋白质与线粒体的可能关联，首先使用免疫定位试验在Hep 3B和人肺动脉内皮细胞和平滑肌细胞。线粒体的强抗STAT 6免疫定位在首先用洋地黄皂苷-蔗糖缓冲液洗涤以去除大量可溶性STAT蛋白的细胞的荧光和电子显微镜测定中是明显的。在活细胞成像研究中，STAT 6-GFP，而不是N1-GFP，被观察到组成型共定位与MitoTracker-和四甲基罗丹明乙酯（TMRE）-阳性线粒体，并与线粒体F1-ATP酶固定后通过免疫荧光测定。这种关联是Tyr磷酸化独立的，因为缺少SH 2结构域（517-632）和甘氨酸激活的Y 641磷酸化位点的STAT 6截短蛋白（STAT 61 -459-GFP）也在MitoTracker阳性线粒体中积累。这与意外发现一致，即抗STAT 6免疫荧光也与来自野生型和STAT 6SH 2-/SH 2-小鼠的小鼠胚胎成纤维细胞（MEF）中的线粒体相关。来自后一种小鼠的MEF（其在1996年被工程化以在STAT 6 SH 2结构域（氨基酸505-584）中缺失）表达在全细胞和富集的部分中可检测到的免疫特异性150 kDa蛋白。总之，目前的数据提供了第一个明确的证据，任何STAT蛋白家族成员与线粒体的关联-STAT 6。
The transcription factor STAT3 has been previously reported to be associated with mitochondria. However, we have been unable to visualize an association of STAT3-GFP, STAT3-DsRed or STAT3-Flag with mitochondria in human Hep3B hepatocytes thus far even though an association of these molecules with other cytoplasmic organelles (endosomes) was readily demonstrable. We then addressed the broader question of a possible association of other STAT-family of proteins with mitochondria by first using immunolocalization assays in Hep3B and human pulmonary arterial endothelial and smooth muscle cells. Strong anti-STAT6-immunolocalization with mitochondria was apparent in fluorescence and electron microscopy assays of cells first washed with a digitonin-sucrose buffer to remove bulk soluble STAT proteins. In live-cell imaging studies, STAT6-GFP, but not N1-GFP, was observed to constitutively colocalize with MitoTracker- and tetramethylrhodamine ethyl ester (TMRE)-positive mitochondria, and with mitochondrial F1-ATPase when assayed by immunofluorescence after fixation. This association was Tyr-phosphorylation independent in that a STAT6 truncated protein (STAT61-459-GFP) which lacked the SH2 domain (517–632) and the cytokine-activated Y641 phosphorylation site also accumulated in MitoTracker-positive mitochondria. This was consistent with the unexpected discovery that anti-STAT6-immunofluoresence also associated with mitochondria in mouse embryo fibroblasts (MEFs) from both wild-type and the STAT6SH2-/SH2- mouse. MEFs from the latter mouse, which had been engineered in 1996 to be deleted in the STAT6 SH2 domain (amino acids 505–584) expressed an immune-specific ∼50 kDa protein detectable in whole cell and mitochondria-enriched fractions. Taken together, the present data provide the first definitive evidence of the association of any STAT-protein family member with mitochondria - that of STAT6.
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DOI：
10.1126/science.1164551
发表时间：
2009-02-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Wegrzyn J;Potla R;Chwae YJ;Sepuri NB;Zhang Q;Koeck T;Derecka M;Szczepanek K;Szelag M;Gornicka A;Moh A;Moghaddas S;Chen Q;Bobbili S;Cichy J;Dulak J;Baker DP;Wolfman A;Stuehr D;Hassan MO;Fu XY;Avadhani N;Drake JI;Fawcett P;Lesnefsky EJ;Larner AC
通讯作者：
Larner AC
影响因子：
9.5
作者：
Boengler K;Hilfiker-Kleiner D;Heusch G;Schulz R
通讯作者：
Schulz R
DOI：
10.1016/j.bbrc.2010.10.112
发表时间：
2010-11-26
影响因子：
3.1
作者：
Chueh FY;Leong KF;Yu CL
通讯作者：
Yu CL
影响因子：
16
作者：
Chen, Liuh-Yow;Zhang, Yi;Zhang, Qinfen;Li, Hongzhi;Luo, Zhenhua;Fang, Hezhi;Kim, Sok Ho;Qin, Li;Yotnda, Patricia;Xu, Jianming;Tu, Benjamin P.;Bai, Yidong;Zhou Songyang
通讯作者：
Zhou Songyang
影响因子：
12.8
作者：
Jeong, Kyuho;Kwon, Hayeong;Pak, Yunbae
通讯作者：
Pak, Yunbae