Protein Kinase D1 Mediates Anchorage-dependent and -independent Growth of Tumor Cells via the Zinc Finger Transcription Factor Snail1
Protein Kinase D1 Mediates Anchorage-dependent and -independent Growth of Tumor Cells via the Zinc Finger Transcription Factor Snail1
复制标题
DOI：
10.1074/jbc.m112.370999
复制
发表时间：
2012-09-21
影响因子：
4.8
通讯作者：
Seufferlein, Thomas
中科院分区：
文献类型：
--
作者：
Eiseler, Tim;Koehler, Conny;Seufferlein, Thomas
在这里，我们确定蛋白激酶D1（PKD 1）作为一个主要的调节锚定依赖性和非依赖性的癌细胞的生长通过转录因子蜗牛1控制。使用FRET，我们证明，PKD 1，但不是PKD 2，有效地与蜗牛1在细胞核中相互作用。PKD 1在Ser-11磷酸化Snail 1。在不同的肿瘤细胞中，使用野生型Snail 1和Ser-11磷酸化位点突变体，Snail 1的核质分布没有变化。无论其磷酸化状态或以下的组成型活性PKD的共表达，蜗牛1主要定位于细胞核。我们还确定了PKD 1介导的Snail 1在肿瘤细胞中转录活性调节的新机制。当Snail 1在Ser-11不磷酸化时，辅助阻遏物组蛋白去乙酰化酶1和2以及赖氨酰氧化酶样蛋白3与Snail 1的相互作用受损，这导致Snail 1相关的组蛋白去乙酰化酶活性降低。此外，赖氨酰氧化酶样蛋白3的表达上调异位PKD 1的表达，这意味着蜗牛1驱动的转录的协同调节。PKD 1的异位表达也上调了细胞增殖标记物如Cyclin D1和Ajuba。因此，Snail 1及其Ser-11磷酸化是控制PKD 1介导的不同肿瘤细胞的锚定非依赖性生长和锚定依赖性增殖所必需的。总之，我们的数据表明PKD 1通过Snail 1支持肿瘤细胞的生长至关重要。
We here identify protein kinase D1 (PKD1) as a major regulator of anchorage-dependent and -independent growth of cancer cells controlled via the transcription factor Snail1. Using FRET, we demonstrate that PKD1, but not PKD2, efficiently interacts with Snail1 in nuclei. PKD1 phosphorylates Snail1 at Ser-11. There was no change in the nucleocytoplasmic distribution of Snail1 using wild type Snail1 and Ser-11 phosphosite mutants in different tumor cells. Regardless of its phosphorylation status or following co-expression of constitutively active PKD, Snail1 was predominantly localized to cell nuclei. We also identify a novel mechanism of PKD1-mediated regulation of Snail1 transcriptional activity in tumor cells. The interaction of the co-repressors histone deacetylases 1 and 2 as well as lysyl oxidase-like protein 3 with Snail1 was impaired when Snail1 was not phosphorylated at Ser-11, which led to reduced Snail1-associated histone deacetylase activity. Additionally, lysyl oxidase-like protein 3 expression was up-regulated by ectopic PKD1 expression, implying a synergistic regulation of Snail1-driven transcription. Ectopic expression of PKD1 also up-regulated proliferation markers such as Cyclin D1 and Ajuba. Accordingly, Snail1 and its phosphorylation at Ser-11 were required and sufficient to control PKD1-mediated anchorage-independent growth and anchorage-dependent proliferation of different tumor cells. In conclusion, our data show that PKD1 is crucial to support growth of tumor cells via Snail1.