Biochemical and Structural Insights into Doublecortin-like Kinase Domain 1

Onisha Patel; Weiwen Dai; Mareike Mentzel; Michael D W Griffin; Juliette Serindoux; Yoann Gay; Stefanie Fischer; Shoukat Sterle; Ashleigh Kropp; Christopher J Burns; Matthias Ernst; Michael Buchert; Isabelle S Lucet

doi:10.1016/j.str.2016.07.008

Biochemical and Structural Insights into Doublecortin-like Kinase Domain 1

Structure. 2016 Sep 6;24(9):1550-61. doi: 10.1016/j.str.2016.07.008. Epub 2016 Aug 18.

Authors

Affiliations

¹ The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia. Electronic address: patel.o@wehi.edu.au.
² The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia.
³ Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC 3052, Australia.
⁴ Olivia Newton-John Cancer Research Institute, School of Cancer Medicine, La Trobe University, Heidelberg, VIC 3084, Australia.
⁵ The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3052, Australia. Electronic address: lucet.i@wehi.edu.au.

PMID: 27545623
DOI: 10.1016/j.str.2016.07.008

Abstract

Doublecortin-like kinase 1 (DCLK1) is a serine/threonine kinase that belongs to the family of microtubule-associated proteins. Originally identified for its role in neurogenesis, DCLK1 has recently been shown to regulate biological processes outside of the CNS. DCLK1 is among the 15 most common putative driver genes for gastric cancers and is highly mutated across various other human cancers. However, our present understanding of how DCLK1 dysfunction leads to tumorigenesis is limited. Here, we provide evidence that DCLK1 kinase activity negatively regulates microtubule polymerization. We present the crystal structure of the DCLK1 kinase domain at 1.7 Å resolution, providing detailed insight into the ATP-binding site that will serve as a framework for future drug design. This structure also allowed for the mapping of cancer-causing mutations within the kinase domain, suggesting that a loss of kinase function may contribute to tumorigenesis.

MeSH terms

Adenosine Triphosphate / chemistry*
Adenosine Triphosphate / metabolism
Amino Acid Motifs
Animals
Antineoplastic Agents / chemistry
Antineoplastic Agents / pharmacology*
Benzodiazepinones / chemistry
Benzodiazepinones / pharmacology
Binding Sites
Crystallography, X-Ray
Doublecortin-Like Kinases
Gene Expression
HEK293 Cells
Humans
Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
Intracellular Signaling Peptides and Proteins / chemistry
Intracellular Signaling Peptides and Proteins / genetics
Intracellular Signaling Peptides and Proteins / metabolism
Mice
Mice, Inbred C57BL
Mice, Transgenic
Microtubules / drug effects
Microtubules / metabolism
Microtubules / ultrastructure
Models, Molecular
Mutation*
Protein Binding
Protein Domains
Protein Kinase Inhibitors / chemistry
Protein Kinase Inhibitors / pharmacology
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Protein Serine-Threonine Kinases / chemistry
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
Protein Structure, Secondary
Pyrimidines / chemistry
Pyrimidines / pharmacology
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Stomach Neoplasms / drug therapy*
Stomach Neoplasms / enzymology
Stomach Neoplasms / genetics
Stomach Neoplasms / pathology
Substrate Specificity
Tubulin Modulators / chemistry
Tubulin Modulators / pharmacology
Xenograft Model Antitumor Assays

Substances

Antineoplastic Agents
Benzodiazepinones
Intracellular Signaling Peptides and Proteins
LRRK2-IN1
NVP-TAE684
Protein Kinase Inhibitors
Pyrimidines
Recombinant Proteins
Tubulin Modulators
XMD 8-92
Adenosine Triphosphate
DCLK1 protein, human
Doublecortin-Like Kinases
Protein Serine-Threonine Kinases