CDK11 negatively regulates Wnt/β-catenin signaling in the endosomal compartment by affecting microtubule stability

Danmin Ou; Lin Chen; Jiang He; Zhuoxian Rong; Jie Gao; Zhi Li; Liyu Liu; Feiyu Tang; Jiang Li; Yuezhen Deng; Lunquan Sun

doi:10.20892/j.issn.2095-3941.2019.0229

CDK11 negatively regulates Wnt/β-catenin signaling in the endosomal compartment by affecting microtubule stability

Cancer Biol Med. 2020 May 15;17(2):328-342. doi: 10.20892/j.issn.2095-3941.2019.0229.

Authors

Danmin Ou¹, Lin Chen¹, Jiang He¹, Zhuoxian Rong¹, Jie Gao¹, Zhi Li^{1

2

3}, Liyu Liu¹, Feiyu Tang¹, Jiang Li¹, Yuezhen Deng^{1

2

3}, Lunquan Sun^{1

2

3

4}

Affiliations

¹ Department of Oncology, Center for Molecular Medicine, Xiangya Hospital, Central South University, Changsha 410008, China.
² International Cooperation Base of Cancer Precision Therapy, Department of Science and Technology of Hunan Province, Changsha 410008, China.
³ Key Laboratory of Molecular Radiation Oncology of Hunan Province, Changsha 410008, China.
⁴ National Clinical Research Center for Geriatric Disorders, Changsha 410008, China.

PMID: 32587772
PMCID: PMC7309457
DOI: 10.20892/j.issn.2095-3941.2019.0229

Abstract

Objectives: Improper activation of Wnt/β-catenin signaling has been implicated in human diseases. Beyond the well-studied glycogen synthase kinase 3β (GSK3β) and casein kinase 1 (CK1), other kinases affecting Wnt/β-catenin signaling remain to be defined. Methods:To identify the kinases that modulate Wnt/β-catenin signaling, we applied a kinase small interfering RNA (siRNA) library screen approach. Luciferase assays, immunoblotting, and real-time polymerase chain reaction (PCR) were performed to confirm the regulation of the Wnt/β-catenin signaling pathway by cyclin-dependent kinase 11 (CDK11) and to investigate the underlying mechanism. Confocal immunofluorescence, coimmunoprecipitation (co-IP), and scratch wound assays were used to demonstrate colocalization, detect protein interactions, and explore the function of CDK11. Results: CDK11 was found to be a significant candidate kinase participating in the negative control of Wnt/β-catenin signaling. Down-regulation of CDK11 led to the accumulation of Wnt/β-catenin signaling receptor complexes, in a manner dependent on intact adenomatosis polyposis coli (APC) protein. Further analysis showed that CDK11 modulation of Wnt/β-catenin signaling engaged the endolysosomal machinery, and CDK11 knockdown enhanced the colocalization of Wnt/β-catenin signaling receptor complexes with early endosomes and decreased colocalization with lysosomes. Mechanistically, CDK11 was found to function in Wnt/β-catenin signaling by regulating microtubule stability. Depletion of CDK11 down-regulated acetyl-α-tubulin. Moreover, co-IP assays demonstrated that CDK11 interacts with the α-tubulin deacetylase SIRT2, whereas SIRT2 down-regulation in CDK11-depleted cells reversed the accumulation of Wnt/β-catenin signaling receptor complexes. CDK11 was found to suppress cell migration through altered Wnt/β-catenin signaling. Conclusions: CDK11 is a negative modulator of Wnt/β-catenin signaling that stabilizes microtubules, thus resulting in the dysregulation of receptor complex trafficking from early endosomes to lysosomes.

Keywords: CDK11; SIRT2; Wnt/β-catenin signaling; endosome; microtubule.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenomatous Polyposis Coli Protein
Cell Line, Tumor
Cyclin-Dependent Kinases / genetics
Cyclin-Dependent Kinases / metabolism*
Down-Regulation*
Endosomes / genetics
Endosomes / metabolism*
Humans
Luciferases / analysis
Luciferases / genetics
Microtubules / genetics
Microtubules / metabolism*
Sirtuin 2 / genetics
Sirtuin 2 / metabolism
Wnt Proteins
Wnt Signaling Pathway
beta Catenin

Substances

Adenomatous Polyposis Coli Protein
Wnt Proteins
beta Catenin
Luciferases
CDK19 protein, human
Cyclin-Dependent Kinases
SIRT2 protein, human
Sirtuin 2