Phosphorylation of ERK5 on Thr732 is associated with ERK5 nuclear localization and ERK5-dependent transcription.
Phosphorylation of ERK5 on Thr732 is associated with ERK5 nuclear localization and ERK5-dependent transcription.
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DOI：
10.1371/journal.pone.0117914
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Nakahata N
中科院分区：
文献类型：
--
作者：
Honda T;Obara Y;Yamauchi A;Couvillon AD;Mason JJ;Ishii K;Nakahata N
细胞外信号调节激酶（ERK）在许多细胞过程中发挥关键作用，包括增殖和分化。ERK 5在N-末端含有激酶结构域，并且独特的延伸C-末端包括增强ERK 5依赖性转录的多个自磷酸化位点。然而，磷酸化在不同位点的影响仍不清楚。在这项研究中，我们研究了磷酸化在ERK 5 C-末端的作用。我们发现，一个组成型活性MEK 5突变体磷酸化ERK 5在TEY基序，导致连续的多个C-末端残基，包括Thr 732和Ser 769/773/775的自磷酸化。然而，当ERK 1/2被致癌RAS突变体选择性激活时，ERK 5在Thr 732的磷酸化被诱导，而不影响TEY或Ser 769/773/775的磷酸化状态。Thr 732磷酸化是U 0126敏感的，并且在ERK 5的激酶死亡突变体中也观察到，这表明ERK 1/2可以在Thr 732磷酸化ERK 5。在HEK 293和PC 12细胞中，表皮生长因子和神经生长因子也分别促进这种磷酸化。ERK 5-T732 A突变体在基础条件下定位于胞质溶胶中。相反，通过RAS-ERK 1/2途径在Thr 732处磷酸化的ERK 5和模拟磷酸化形式的ERK 5-T732 E定位于细胞核和胞质溶胶中。最后，ER-32A和U 0126阻断ERK 5依赖性MEF 2C转录活性。基于这些发现，我们提出了一种新的串扰机制，其中ERK 1/2，激活生长因子刺激，磷酸化ERK 5在Thr 732。该磷酸化事件负责ERK 5核定位和ERK 5依赖性转录。
Extracellular signal-regulated kinases (ERKs) play critical roles in numerous cellular processes, including proliferation and differentiation. ERK5 contains a kinase domain at the N-terminal, and the unique extended C-terminal includes multiple autophosphorylation sites that enhance ERK5-dependent transcription. However, the impact of phosphorylation at the various sites remain unclear. In this study, we examined the role of phosphorylation at the ERK5 C-terminal. We found that a constitutively active MEK5 mutant phosphorylated ERK5 at the TEY motif, resulting in the sequential autophosphorylation of multiple C-terminal residues, including Thr732 and Ser769/773/775. However, when ERK1/2 was selectively activated by an oncogenic RAS mutant, ERK5 phosphorylation at Thr732 was induced without affecting the phosphorylation status at TEY or Ser769/773/775. The Thr732 phosphorylation was U0126-sensitive and was observed in a kinase-dead mutant of ERK5 as well, suggesting that ERK1/2 can phosphorylate ERK5 at Thr732. This phosphorylation was also promoted by epidermal growth factor and nerve growth factor in HEK293 and PC12 cells, respectively. The ERK5–T732A mutant was localized in the cytosol under basal conditions. In contrast, ERK5 phosphorylated at Thr732 via the RAS-ERK1/2 pathway and ERK5–T732E, which mimics the phosphorylated form, were localized in both the nucleus and cytosol. Finally, ER–32A and U0126 blocked ERK5-dependent MEF2C transcriptional activity. Based on these findings, we propose a novel cross-talk mechanism in which ERK1/2, following activation by growth factor stimulation, phosphorylates ERK5 at Thr732. This phosphorylation event is responsible for ERK5 nuclear localization and ERK5-dependent transcription.
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影响因子：
4.8
作者：
Kamakura, S;Moriguchi, T;Nishida, E
通讯作者：
Nishida, E
DOI：
10.1152/ajprenal.00130.2007
发表时间：
2007-11-01
影响因子：
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作者：
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Kato, Y;Kravchenko, VV;Lee, JD
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影响因子：
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作者：
Obara, Yutaro;Okano, Yumiko;Nakahata, Norimichi
通讯作者：
Nakahata, Norimichi
影响因子：
4.8
作者：
Buschbeck, M;Ullrich, A
通讯作者：
Ullrich, A