Persulfidation of transcription factor FOXO1 at cysteine 457: A novel mechanism by which H(2)S inhibits vascular smooth muscle cell proliferation.
Persulfidation of transcription factor FOXO1 at cysteine 457: A novel mechanism by which H(2)S inhibits vascular smooth muscle cell proliferation.
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转录因子 FOXO1 在半胱氨酸 457 处的过硫化：H2S 抑制血管平滑肌细胞增殖的新机制
DOI：
10.1016/j.jare.2020.06.023
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发表时间：
2021-01
影响因子：
10.7
通讯作者：
Jin H
中科院分区：
文献类型：
--
作者：
Tian X;Zhou D;Zhang Y;Song Y;Zhang Q;Bu D;Sun Y;Wu L;Long Y;Tang C;Du J;Huang Y;Jin H
H2S抑制以fox01为靶点的VSMC增殖机制示意图。H2S在Cys457位点过硫化FOXO1，降低FOXO1 Ser256位点的磷酸化，阻止FOXO1细胞核排斥到细胞质，进一步保护et -1诱导的VSMC增殖。FOXO1参与H2S对血管平滑肌细胞增殖的抑制作用。H2S通过维持FOXO1活性抑制血管平滑肌细胞增殖。H2S通过过硫化保持FOXO1活性。H2S使FOXO1在Cys457位点过硫，随后阻止FOXO1在Ser256位点磷酸化。结果为抗血管重构的治疗策略提供了新的思路。血管平滑肌细胞（VSMCs）的增殖是许多心血管疾病发生的重要生理病理基础。内源性硫化氢（H2S）是第三种气体递质，通过抑制VSMC增殖来保护血管结构。然而，H2S抑制VSMC增殖的机制尚不完全清楚。本研究旨在探讨H2S过硫化转录因子FOXO1是否抑制VSMC增殖。内皮素-1 （ET-1）诱导VSMC A7r5细胞增殖后，采用Western blotting检测FOXO1磷酸化水平和增殖细胞核抗原（PCNA）表达，免疫荧光检测FOXO1核排斥程度和细胞核内PCNA荧光信号，生物素开关法检测FOXO1过硫化程度。结果表明，ET-1刺激增加了细胞增殖，使FOXO1磷酸化，使FOXO1核排除到细胞质中。然而，H2S供体NaHS预处理成功地消除了et -1诱导的VSMC增殖、FOXO1磷酸化和细胞质中FOXO1核排斥的增加。从机制上讲，H2S使A7r5和293T细胞中的fox01蛋白过硫化，硫醇还原剂DTT逆转了这一作用。此外，FOXO1的C457S突变消除了h2s诱导的FOXO1在细胞中的过硫化作用以及随后对FOXO1 Ser256位点磷酸化、FOXO1核排斥对细胞质和细胞增殖的抑制作用。因此，我们的研究结果表明H2S可能通过使FOXO1在Cys457位点过硫并随后阻止FOXO1在Ser256位点磷酸化来抑制VSMC增殖。
Schematic diagram of the mechanism by which H2S inhibits VSMC proliferation targeting on the FOXO1. H2S persulfidates FOXO1 at Cys457, which reduces FOXO1 phosphorylation at Ser256, prevents the FOXO1 nucleus exclusion to the cytoplasm, and further protecting against ET-1-induced VSMC proliferation. FOXO1 is involved in the inhibitory effect of H2S on vascular smooth muscle cell proliferation. H2S inhibits vascular smooth muscle cell proliferation by maintaining FOXO1 activity. H2S preserves FOXO1 activity by persulfidation. H2S persulfidates FOXO1 at Cys457 and subsequently prevents FOXO1 phosphorylation at Ser256. The results provide new ideas for therapeutic strategies for anti-vascular remodeling. The proliferation of vascular smooth muscle cells (VSMCs) is an important physiological and pathological basis for many cardiovascular diseases. Endogenous hydrogen sulfide (H2S), the third gasotransmitter, is found to preserve vascular structure by inhibiting VSMC proliferation. However, the mechanism by which H2S suppresses VSMC proliferation has not been fully clear. This study aimed to explore whether H2S persulfidates the transcription factor FOXO1 to inhibit VSMC proliferation. After the proliferation of VSMC A7r5 cells was induced by endothelin-1 (ET-1), FOXO1 phosphorylation and proliferating cell nuclear antigen (PCNA) expression were detected by Western blotting, the degree of FOXO1 nuclear exclusion and PCNA fluorescent signals in the nucleus were detected by immunofluorescence, and the persulfidation of FOXO1 was measured through a biotin switch assay. The results showed that ET-1 stimulation increased cell proliferation, FOXO1 phosphorylation and FOXO1 nuclear exclusion to the cytoplasm in the cells. However, pretreatment with NaHS, an H2S donor, successfully abolished the ET-1-induced increases in the VSMC proliferation, FOXO1 phosphorylation, and FOXO1 nuclear exclusion to the cytoplasm. Mechanistically, H2S persulfidated the FOXO1 protein in A7r5 and 293T cells, and the thiol reductant DTT reversed this effect. Furthermore, the C457S mutation of FOXO1 abolished the H2S-induced persulfidation of FOXO1 in the cells and the subsequent inhibitory effects on FOXO1 phosphorylation at Ser256, FOXO1 nuclear exclusion to the cytoplasm and cell proliferation. Thus, our findings demonstrated that H2S might inhibit VSMC proliferation by persulfidating FOXO1 at Cys457 and subsequently preventing FOXO1 phosphorylation at Ser256.
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DOI：
10.1126/science.1162667
发表时间：
2008-10-24
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Yang G;Wu L;Jiang B;Yang W;Qi J;Cao K;Meng Q;Mustafa AK;Mu W;Zhang S;Snyder SH;Wang R
通讯作者：
Wang R
通过 Ca(2)/钙调蛋白依赖性蛋白激酶 II 的 S-硫化作用改善心力衰竭中的线粒体功能障碍。
影响因子：
11.4
作者：
Wu D;Hu Q;Tan B;Rose P;Zhu D;Zhu YZ
通讯作者：
Zhu YZ
影响因子：
56.9
作者：
Tran, H;Brunet, A;Greenberg, ME
通讯作者：
Greenberg, ME
影响因子：
4.8
作者：
Majumder, Suravi;Ren, Lu;Sen, Utpal
通讯作者：
Sen, Utpal
影响因子：
4.8
作者：
Kim, Sunhong;Kim, Yongsung;Chung, Jongkyeong
通讯作者：
Chung, Jongkyeong