Requirement for protein kinase A in the phosphorylation of the TGFβ receptor-interacting protein km23-1 as a component of TGFβ downstream effects.
Requirement for protein kinase A in the phosphorylation of the TGFβ receptor-interacting protein km23-1 as a component of TGFβ downstream effects.
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DOI：
10.1016/j.yexcr.2012.12.029
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发表时间：
2013-04-01
影响因子：
3.7
通讯作者：
Mulder, Kathleen M.
中科院分区：
文献类型：
--
作者：
Jin, Qunyan;Zhong, Yan;Mulder, Kathleen M.
km23-1先前被鉴定为TGFß-受体相互作用蛋白，在TGFß刺激后在丝氨酸上磷酸化。在本报告中，我们研究了km23-1磷酸化在TGFß/蛋白激酶A （PKA）信号传导的下游效应中的作用。利用磷酸化位点预测软件，我们发现km23-1有两个潜在的PKA共识磷酸化位点。体外激酶实验进一步表明，PKA直接磷酸化km23-1的丝氨酸73 （S73）。此外，我们的研究结果表明，pka特异性抑制剂H89在TGFß刺激后减少了km23-1对S73的磷酸化。综上所述，我们的研究结果表明TGFß诱导PKA活性导致S73上km23-1的磷酸化。为了评估TGFß刺激后PKA磷酸化km23-1对S73 （S73-km23-1）的机制，我们进行了免疫沉淀(IP)/blot分析，结果表明TGFß在体内调节PKA调控亚基RIß和km23-1之间的复合物形成。此外，km23-1的S73A突变体（S73A-km23-1）不能被PKA磷酸化，抑制TGFß诱导km23-1-dynein复合物和激活素响应元件（activin-responsive element， ARE）的转录激活。此外，我们的研究结果表明，km23-1是TGFß激活cAMP-responsive element （CRE）转录所必需的，而S73-km23-1是CRE依赖性TGFß刺激纤维连接蛋白（FN）转录所必需的。总的来说，我们的研究结果首次证明了TGFß/PKA在S73上磷酸化km23-1是ARE和cre介导的下游事件（包括FN诱导）所必需的。
km23-1 was previously identified as a TGFß-receptor interacting protein that was phosphorylated on serines after TGFß stimulation. In the current report, we examined the role of km23-1 phosphorylation in the downstream effects of TGFß/ protein kinase A (PKA) signaling. Using phosphorylation site prediction software, we found that km23-1 has two potential PKA consensus phosphorylation sites. In vitro kinase assays further demonstrated that PKA directly phosphorylates km23-1 on serine 73 (S73). Moreover, our results show that the PKA-specific inhibitor H89 diminishes phosphorylation of km23-1 on S73 after TGFß stimulation. Taken together, our results demonstrate that TGFß induction of PKA activity results in phosphorylation of km23-1 on S73. In order to assess the mechanisms underlying PKA phosphorylation of km23-1 on S73 (S73-km23-1) after TGFß stimulation, immunoprecipitation (IP)/blot analyses were performed, which demonstrate that TGFß regulates complex formation between the PKA regulatory subunit RIß and km23-1 in vivo. In addition, an S73A mutant of km23-1 (S73A-km23-1), which could not be phosphorylated by PKA, inhibited TGFß induction of the km23-1-dynein complex and transcriptional activation of the activin-responsive element (ARE). Furthermore, our results show that km23-1 is required for cAMP-responsive element (CRE) transcriptional activation by TGFß, with S73-km23-1 being required for the CRE-dependent TGFß stimulation of fibronectin (FN) transcription. Collectively, our results demonstrate for the first time that TGFß/PKA phosphorylation of km23-1 on S73 is required for ARE- and CRE-mediated downstream events that include FN induction.
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影响因子：
3.7
作者：
Chowdhury S;Howell GM;Rajput A;Teggart CA;Brattain LE;Weber HR;Chowdhury A;Brattain MG
通讯作者：
Brattain MG
影响因子：
3.5
作者：
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通讯作者：
Zhao, SY
影响因子：
4.8
作者：
Hocevar, BA;Prunier, C;Howe, PH
通讯作者：
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影响因子：
4.8
作者：
Jin, Qunyan;Ding, Wei;Mulder, Kathleen M.
通讯作者：
Mulder, Kathleen M.
DOI：
10.1016/j.bbrc.2006.07.057
发表时间：
2006-09-15
影响因子：
3.1
作者：
Lai, Lingyun;Chen, Jing;Gu, Yong
通讯作者：
Gu, Yong