Prohibitin, STAT3 and SH2D4A physically and functionally interact in tumor cell mitochondria.
Prohibitin, STAT3 and SH2D4A physically and functionally interact in tumor cell mitochondria.
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DOI：
10.1038/s41419-020-03220-3
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发表时间：
2020-11-30
影响因子：
9
通讯作者：
Roessler S
中科院分区：
文献类型：
--
作者：
Ploeger C;Huth T;Sugiyanto RN;Pusch S;Goeppert B;Singer S;Tabti R;Hausser I;Schirmacher P;Désaubry L;Roessler S
8p染色体在各种癌症实体中经常被删除，并且已被证明与患者生存率低有关。SH2D4A位于染色体8p上，可阻止促肿瘤转录因子STAT3的核易位。在这里，我们研究了SH2D4A和STAT3的相互作用，以揭示STAT3与肿瘤抑制因子SH2D4A合作的非规范功能。使用免疫沉淀-质谱（IP-MS）方法，我们鉴定了线粒体支架蛋白禁止蛋白1 （PHB1）和禁止蛋白2 （PHB2）以及其他可能与SH2D4A结合的蛋白。共免疫沉淀和邻近连接实验证实了STAT3、PHB1和SH2D4A在原位和体外的直接相互作用。此外，细胞分离和免疫荧光染色显示这些蛋白与线粒体共定位。这些相互作用被小分子和PHB配体FL3选择性地中断。此外，在二甲基氧基草酸甘氨酸（DMOG）处理下，FL3导致STAT3蛋白水平降低，STAT3转录活性降低，HIF1α蛋白稳定。此外，线粒体融合和裂变标志物L-OPA1、Mfn1和FIS1在FL3处理后出现异常。这种失调的形态伴随着线粒体呼吸的显著减少，因此，FL3显著降低了线粒体呼吸能力。相反，敲除SH2D4A增加了线粒体呼吸，而FL3逆转了敲除SH2D4A的作用。上述结果表明，SH2D4A和PHB1的相互作用参与线粒体功能和完整性。已证实的与STAT3的相互作用，伴随着转录活性的降低，进一步表明SH2D4A将STAT3与其线粒体功能联系起来，抑制phb相互作用可能对STAT3激活的肿瘤细胞有治疗作用。
Chromosome 8p is frequently deleted in various cancer entities and has been shown to correlate with poor patient survival. SH2D4A is located on chromosome 8p and prevents the nuclear translocation of the pro-tumorigenic transcription factor STAT3. Here, we investigated the interaction of SH2D4A and STAT3 to shed light on the non-canonical functions of STAT3 in cooperation with the tumor suppressor SH2D4A. Using an immunoprecipitation-mass spectrometry (IP-MS) approach, we identified the mitochondrial scaffold proteins prohibitin 1 (PHB1) and prohibitin 2 (PHB2) among other proteins to potentially bind to SH2D4A. Co-immunoprecipitation and proximity ligation assays confirmed direct interactions of STAT3, PHB1, and SH2D4A in situ and in vitro. In addition, cell fractionation and immunofluorescence staining revealed co-localization of these proteins with mitochondria. These interactions were selectively interrupted by the small molecule and PHB ligand FL3. Furthermore, FL3 led to a reduction of STAT3 protein levels, STAT3 transcriptional activity, and HIF1α protein stabilization upon dimethyloxalylglycine (DMOG) treatment. Besides, mitochondrial fusion and fission markers, L-OPA1, Mfn1, and FIS1, were dysregulated upon FL3 treatment. This dysregulated morphology was accompanied by significant reduction of mitochondrial respiration, thus, FL3 significantly diminished mitochondrial respirational capacity. In contrast, SH2D4A knockout increased mitochondrial respiration, whereas FL3 reversed the effect of SH2D4A knockout. The here described results indicate that the interaction of SH2D4A and PHB1 is involved in the mitochondrial function and integrity. The demonstrated interaction with STAT3, accompanied by its reduction of transcriptional activity, further suggests that SH2D4A is linking STAT3 to its mitochondrial functions, and inhibition of PHB-interaction may have therapeutic effects in tumor cells with STAT3 activation.
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影响因子：
46.9
作者：
Doench JG;Fusi N;Sullender M;Hegde M;Vaimberg EW;Donovan KF;Smith I;Tothova Z;Wilen C;Orchard R;Virgin HW;Listgarten J;Root DE
通讯作者：
Root DE
影响因子：
3.8
作者：
Garama, Daniel J.;White, Christine L.;Gough, Daniel J.
通讯作者：
Gough, Daniel J.
影响因子：
--
作者：
Kluth M;Jung S;Habib O;Eshagzaiy M;Heinl A;Amschler N;Masser S;Mader M;Runte F;Barow P;Frogh S;Omari J;Möller-Koop C;Hube-Magg C;Weischenfeldt J;Korbel J;Steurer S;Krech T;Huland H;Graefen M;Minner S;Sauter G;Schlomm T;Simon R
通讯作者：
Simon R
影响因子：
4.8
作者：
Fusaro, G;Dasgupta, P;Chellappan, S
通讯作者：
Chellappan, S
影响因子：
11.2
作者：
Kathiria AS;Neumann WL;Rhees J;Hotchkiss E;Cheng Y;Genta RM;Meltzer SJ;Souza RF;Theiss AL
通讯作者：
Theiss AL