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Published on: March 5, 2019
Chemical Biology Toolkit for DCLK1 Reveals Connection to RNA Processing
Yan Liu1, Fleur M Ferguson2, Lianbo Li1
1Department 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.
Abstract:
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.
Insights
Doublecortin-like kinase 1 (DCLK1) is crucial for brain development and cancer. Researchers developed a toolkit to study DCLK1's role in cancer, revealing its link to RNA processing and identifying CDK11 as a potential substrate.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Doublecortin-like kinase 1 (DCLK1) plays a vital role in neurogenesis.
- DCLK1 overexpression is linked to poor prognosis in various cancers.
- The precise functions and context-dependent roles of DCLK1 in cancer remain largely unknown.
Purpose of the Study:
- To develop and utilize a toolkit for investigating DCLK1 signaling pathways in cancer.
- To explore the association between DCLK1 kinase activity and cancer-related biological processes.
- To identify novel DCLK1 substrates and downstream pathways.
Main Methods:
- Development of a DCLK1 inhibitor (DCLK1-IN-1) and complementary resistant/dead mutants (G532A, D511N, D533N).
- Engineering a DCLK1-dependent cancer cell line for functional studies.
- Application of phosphoproteomics to identify DCLK1 substrates.
Main Results:
- The developed toolkit successfully enabled the investigation of DCLK1 kinase activity in a cancer cell line.
- A significant association was found between DCLK1 activity and RNA processing.
- CDK11 was identified as a potential substrate of DCLK1 through phosphoproteomic analysis.
Conclusions:
- The DCLK1 toolkit is effective for dissecting DCLK1-regulated signaling in cancer.
- DCLK1 kinase activity is implicated in RNA processing pathways.
- Further research into DCLK1 and its substrate CDK11 may reveal new therapeutic strategies for cancer.
