Cell fate signalling by the DYRK protein kinases, relatives of the MAP kinases

双特异性酪氨酸磷酸化调节蛋白激酶(DYRKs)是一个与MAP相关的5个蛋白激酶家族(如ERK)。DYRK似乎在正常发育和内环境稳定中很重要，在某些疾病(唐斯综合征、癌症)中可能会被解除调控，但它们的确切功能仍然是个谜。基于我们在MAPK生物学上的成功(1)，我们对DYRK及其在控制细胞周期、存活、分化和自噬中的作用越来越感兴趣。然而，很少有Dyrk底物或Dyrk靶基因被定义来解释这些酶如何调控这种细胞命运的决定。我们现在已经建立了一组细胞系，它们展示了Tet调控的不同Dyrk蛋白激酶的表达。此外，我们还有一个细胞小组，其中单个DYRK已经被CRISPR/Cas9技术敲除，并可以获得独特的、专有的Dyrk抑制剂。我们正在使用这个‘Dyrk工具包’来研究Dyrk激活/失活的细胞后果，并应用转录组学/磷酸蛋白质组学方法来了解DYRK的生物学功能。例如，我们已经使用这个资源展示了DYRK1B和DYRK1A使细胞周期蛋白D1磷酸化来控制细胞周期的G1期，并且最近使用质谱仪来识别和验证全新的DYRK1B底物(未发表的观察结果)。在这个项目中，学生将使用‘Dyrk工具包’来研究单个DYRK如何影响细胞周期、细胞存活和自噬，包括识别新的Dyrk底物。该项目将生成并整合基因组和蛋白质组学数据，以确定Dyrk蛋白激酶的生物学功能。

The dual-specificity tyrosine-phosphorylation-regulated kinases (DYRKs) are a family of five protein kinases related to the MAP kinases (e.g. ERK). The DYRKs appear to be important in normal development and homeostasis and may be de-regulated in certain diseases (Downs Syndrome, Cancer) but their precise functions remain enigmatic. Based on our success in MAPK biology (1) we are increasingly interested in the DYRKs and their role in control of cell cycle, survival, differentiation and autophagy. However, few DYRK substrates or DYRK target genes have been defined to explain how these enzymes regulate such cell fate decisions.We have now established a panel of cell lines that exhibit Tet-regulated expression of the different DYRK protein kinases. In addition we have a panel of cells in which the individual DYRKs have been knocked out by CRISPR/Cas9 technology and have access to unique, proprietary DYRK inhibitors. We are using this 'DYRK toolkit' to investigate the cellular consequences of DYRK activation/inactivation and to apply transcriptomics/phosphoproteomics approaches to understand the biological functions of the DYRKs. For example, we have used this resource to show that DYRK1B and DYRK1A phosphorylate cyclin D1 to control the G1 phase of the cell cycle and have used recently mass spectrometry to identify and validate completely new DYRK1B substrates (unpublished observations).In this project the student will use the 'DYRK toolkit' to investigate how individual DYRKs influence cell cycle, cell survival and autophagy including the identification of new DYRK substrates. The project will generate and integrate genomic and proteomics data to define the biological functions of the DYRK protein kinases.