Smurf1 regulates ameloblast polarization by ubiquitination-mediated degradation of RhoA.
Smurf1 regulates ameloblast polarization by ubiquitination-mediated degradation of RhoA.
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DOI：
10.1111/cpr.13387
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发表时间：
2023-04
影响因子：
8.5
通讯作者：
中科院分区：
文献类型：
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细胞极性对成釉细胞分化和成釉细胞的形成至关重要。Smurf 1可以通过泛素化降解特定底物来介导细胞极化。但目前尚不清楚Smurf 1是否可以调节成釉细胞的极性和潜在的机制。在此，应用免疫荧光染色和RT-qPCR检测Smurf 1和F-actin的表达。建立小鼠下切牙缺损模型。通过扫描电镜、大鼠下切牙培养、Western blot、伤口愈合实验和跨孔迁移实验检测Smurf 1基因敲低对成釉细胞的影响。采用IF双染色、Western blot和免疫共沉淀法检测Smurf 1与RhoA的相互作用。还进行了体内实验。我们发现Smurf 1主要在成釉细胞的细胞膜和细胞皮质中表达，类似于F‐actin。Smurf 1表达沿着成釉细胞极化和分化而增加。敲除Smurf 1后，细胞骨架和细胞形态发生改变，细胞极性受损。Smurf 1通过泛素化降解活化的RhoA调控成釉细胞极性Smurf 1在大鼠下切牙成釉细胞中的局部敲低导致成釉细胞极性丧失、釉基质分泌紊乱和白垩釉，但RhoA抑制剂Y-27632可以逆转这种效应。Smurf 1可通过泛素化降解活化的RhoA来调控成釉细胞的极化，为牙齿发育的研究提供了新的思路。Smurf 1在成釉器和下切牙的牙上皮中呈阳性表达，其表达水平沿着成釉细胞分化成熟而增加。在HAT-7细胞中敲除Smurf 1导致Smurf 1表达定位改变、细胞形态改变和细胞极性丧失。Smurf 1在泛素化-蛋白酶途径中降解活化的RhoA。在体内，Smurf 1基因敲低可导致大鼠下切牙成釉细胞出现白垩釉、成釉细胞极性破坏和釉基质分泌紊乱，但RhoA抑制剂Y-27632可逆转这一效应。
Cell polarity is essential for ameloblast differentiation and enamel formation. Smurf1 can mediate cell polarization through ubiquitination degradation of specific substrates. But it remains unclear whether Smurf1 could regulate ameloblast polarity and the underlying mechanism. Here, immuno‐fluorescence staining and RT‐qPCR were applied to detect the expression of Smurf1 and F‐actin. A mouse lower incisor defect model was constructed. Scanning electron microscope, rat lower incisor culture, western blot, wound healing assay and trans‐well migration assay were performed to detect the influence of Smurf1 knockdown on ameloblast. IF double staining, western blot and co‐immunoprecipitation were conducted to detect the interaction between Smurf1 and RhoA. The in vivo experiment was also performed. We found that Smurf1 was mainly expressed in the membrane and cell cortex of ameloblast, similar to F‐actin. Smurf1 expression increased along ameloblast polarization and differentiation. After knocking down Smurf1, the cytoskeleton and cell morphology changed and the cell polarity was damaged. Smurf1 regulated ameloblast polarity through ubiquitination degradation of activated RhoA in vitro. Local knockdown of Smurf1 in rat lower incisor ameloblast resulted in ameloblast polarity loss, enamel matrix secretion disorder and chalky enamel, but RhoA inhibitor Y‐27632 could reverse this effect. Collectively, Smurf1 could regulate the polarization of ameloblast through ubiquitination degradation of activated RhoA, which contributed to the knowledge of tooth development and provided new research ideas for cell polarity. Smurf1 positively expressed in the dental epithelium of enamel organ and lower incisor, and its expression level increased along the differentiation maturation of ameloblast. Knocking down Smurf1 in HAT‐7 cells resulted in expression location changes of Smurf1, cell morphology alteration and cell polarity loss. Smurf1 degraded activated RhoA in a ubiquitination‐proteinase pathway. In vivo, knockdown of Smurf1 in rat lower incisors' ameloblasts led to chalky enamel, ameloblast polarity disruption and enamel matrix secretion disorder, but RhoA inhibitor Y‐27632 could reverse this effect.
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DOI：
10.1002/jez.b.21254
发表时间：
2009-07-15
影响因子：
2.2
作者：
Boran, Tomas;Peterkova, Renata;Lesot, Herve;Lyons, David B.;Peterka, Miroslav;Klein, Ophir D.
通讯作者：
Klein, Ophir D.
影响因子：
7.7
作者：
Inaba M;Venkei ZG;Yamashita YM
通讯作者：
Yamashita YM
Smurf1 泛素连接酶的 C2 结构域在底物选择中的关键作用
影响因子：
4.8
作者：
Lu, Kefeng;Li, Ping;He, Fuchu
通讯作者：
He, Fuchu
影响因子：
11.2
作者：
Lee, Min-Goo;Jeong, Seong-In;Chi, Sung-Gil
通讯作者：
Chi, Sung-Gil
影响因子：
9.8
作者：
Belyy A;Merino F;Sitsel O;Raunser S
通讯作者：
Raunser S