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Regulation of TGFβ receptor trafficking and signaling by atypical protein kinase C

机译:非典型蛋白激酶C调节TGFβ受体的运输和信号传导

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摘要

Transforming growth factor beta (TGFβ) signaling is linked to the membrane trafficking of TGFβ receptors. The Protein Kinase C (PKC) family of serine/threonine kinases have been implicated in modulating the endocytic processes of various receptors. The present study investigated whether PKC activity plays a role in the trafficking, and signaling of TGFβ receptors, and further explored which PKC isoforms may be responsible for altered TGFβ signaling patterns. Using immunofluorescence microscopy and ~(125)I-TGFβ internalization assays, we show that the pharmacological inhibition of PKC activity alters TGFβ receptor trafficking and delays TGFβ receptor degradation. Consistent with these findings, we demonstrate that PKC inhibition extends TGFβ-dependent Smad2 phosphorylation. Previous studies have shown that PKCζ associates with TGFβ receptors to modulate cell plasticity. We therefore used siRNA directed at the atypical PKC isoforms to investigate if reducing PKCι and PKCζ protein levels would delay TGFβ receptor degradation and extend TGFβ signaling. Our findings suggest that atypical PKC isoforms regulate TGFβ signaling by altering cell surface TGFβ receptor trafficking and degradation.
机译:转化生长因子β(TGFβ)信号转导与TGFβ受体的膜运输有关。丝氨酸/苏氨酸激酶的蛋白激酶C(PKC)家族与调节各种受体的内吞过程有关。本研究调查了PKC活性是否在TGFβ受体的运输和信号传导中起作用,并进一步探讨了哪些PKC亚型可能与TGFβ信号传导模式的改变有关。使用免疫荧光显微镜和〜(125)I-TGFβ内在化分析,我们表明药理学抑制PKC活性改变了TGFβ受体的运输,并延迟了TGFβ受体的降解。与这些发现一致,我们证明PKC抑制作用扩展了TGFβ依赖性的Smad2磷酸化。先前的研究表明,PKCζ与TGFβ受体缔合以调节细胞可塑性。因此,我们使用针对非典型PKC同工型的siRNA来研究降低PKC1和PKCζ蛋白水平是否会延迟TGFβ受体降解并延长TGFβ信号传导。我们的发现表明,非典型PKC亚型通过改变细胞表面TGFβ受体的运输和降解来调节TGFβ信号传导。

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