Department of Biochemistry, The University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390, USA; Department of Radiation Oncology, The University of Texas Southwestern Medical Center at Dallas, Dallas, TX 75390, USA.
Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02215, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
Cell Chem Biol. 2020 Oct 15;27(10):1229-1240.e4. doi: 10.1016/j.chembiol.2020.07.011. Epub 2020 Aug 4.
Doublecortin-like kinase 1 (DCLK1) is critical for neurogenesis, but overexpression is also observed in multiple cancers and is associated with poor prognosis. Nevertheless, the function of DCLK1 in cancer, especially the context-dependent functions, are poorly understood. We present a "toolkit" that includes the DCLK1 inhibitor DCLK1-IN-1, a complementary DCLK1-IN-1-resistant mutation G532A, and kinase dead mutants D511N and D533N, which can be used to investigate signaling pathways regulated by DCLK1. Using a cancer cell line engineered to be DCLK1 dependent for growth and cell migration, we show that this toolkit can be used to discover associations between DCLK1 kinase activity and biological processes. In particular, we show an association between DCLK1 and RNA processing, including the identification of CDK11 as a potential substrate of DCLK1 using phosphoproteomics.
双皮质醇激酶 1（DCLK1）对神经发生至关重要，但也在多种癌症中观察到过表达，并与预后不良相关。然而，DCLK1 在癌症中的功能，特别是上下文相关的功能，仍知之甚少。我们提供了一个“工具包”，其中包括 DCLK1 抑制剂 DCLK1-IN-1、互补的 DCLK1-IN-1 抗性突变 G532A 以及激酶失活突变 D511N 和 D533N，可用于研究由 DCLK1 调控的信号通路。我们使用一种经过工程设计的依赖 DCLK1 生长和细胞迁移的癌细胞系，表明该工具包可用于发现 DCLK1 激酶活性与生物学过程之间的关联。特别是，我们展示了 DCLK1 与 RNA 处理之间的关联，包括使用磷酸化蛋白质组学鉴定 CDK11 作为 DCLK1 的潜在底物。
