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Phosphorylation of MICAL2 by ARG promotes head and neck cancer tumorigenesis by regulating skeletal rearrangement

Oncogene volume 41, pages 334–346 (2022)Cite this article

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Abstract

The actin cytoskeletal architecture provides the structural underpinnings for crucial cellular behaviors. In cancer cells, changes in the actin cytoskeleton may serve as prerequisites for proliferation, invasion, and metastatic dissemination. However, the underlying mechanisms remain largely unknown. Here, we show that MICAL2, which is increased in head and neck squamous cell carcinoma (HNSCC) and inversely associated with patient survival, promotes HNSCC growth, invasion, and migration. MICAL2 serves as a flavoprotein monooxygenase and directly induces actin filament depolymerization by specifically oxidizing the methionine 44 and 47 residues of F-actin. The kinase ARG interacts with MICAL2 and augments MICAL2-mediated actin disassembly. Direct phosphorylation assay and mass spectrometry confirmed that ARG phosphorylates MICAL2 at Tyr445, Tyr463, and Tyr488. Substitution of the Tyr445 or Tyr463 residue of purified recombinant MICAL2-redox with phenylalanine (generating a non-phosphorylatable mutant) abolishes the enhanced MICAL2-mediated F-actin disassembly induced by ARG. Consistently, ectopic expression of non-phosphorylatable MICAL2 mutants (MICAL2Y445F and MICAL2Y463F, not MICAL2Y488F) failed to ameliorate HNSCC cell growth, whereas expression of wild-type MICAL2 or MICAL2Y488F rescued the impaired proliferation induced by MICAL2 knockdown. Moreover, CCG-1423, an inhibitor of MICAL2, was shown to inhibit HNSCC cell proliferation, invasion, and migration. Taken together, our findings indicate that phosphorylation of MICAL2 at Tyr445 and Tyr463 by ARG mediates F-actin disassembly and promotes HNSCC progression.

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Fig. 1: MICAL2 is frequently upregulated in HNSCC, and increased MICAL2 expression is correlated with poor prognosis in patients with HNSCC.
Fig. 2: MICAL2 promotes HNSCC growth both in vitro and in vivo.
Fig. 3: MICAL2 enhances the migration and invasion potential of HNSCC cells and promotes p-EMT.
Fig. 4: ARG, a kinase that interacts with MICAL2, is frequently upregulated in HNSCC and correlated with poor prognosis in patients with HNSCC.
Fig. 5: ARG directly phosphorylates MICAL2 and promotes MICAL2-mediated F-actin disassembly.
Fig. 6: Phosphorylation of Tyr445 and Tyr463 is required for the activity of MICAL2.
Fig. 7: The MICAL2 inhibitor CCG-1423 inhibits cell proliferation, invasion, and migration.

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Acknowledgements

Our sincere appreciation goes out to Professor Ren and Dr. Zhu of the State Key Laboratory of Plant Physiology and Biochemistry of China Agricultural University, for providing technical guidance. We thank the TCGA project for its valuable contributions to HNSCC research. This study was supported by the Natural Science Foundation of China (nos. 81902757, 82073006, and 82002757).

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Author notes

  1. These authors contributed equally: Ze Zhang, Ruoyan Liu, Yafei Wang.

Authors and Affiliations

  1. Department of Maxillofacial and Otorhinolaryngology Oncology and Department of Head and Neck Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, China

    Ze Zhang, Yafei Wang, Yun Wang, Yanjie Shuai, Chuangwu Ke, Rui Jin, Xudong Wang & Jingtao Luo

  2. Key Laboratory of Cancer Prevention and Therapy, Tianjin, China

    Ze Zhang, Ruoyan Liu, Yafei Wang, Yun Wang, Yanjie Shuai, Chuangwu Ke, Rui Jin, Xudong Wang & Jingtao Luo

  3. Tianjin’s Clinical Research Center for Cancer, Tianjin, China

    Ze Zhang, Ruoyan Liu, Yafei Wang, Yun Wang, Yanjie Shuai, Chuangwu Ke, Rui Jin, Xudong Wang & Jingtao Luo

  4. Department of Gynecologic Oncology, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, China

    Ruoyan Liu

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Contributions

Conception and design: ZZ and JL. Development of methodology: ZZ, YW and YW. Acquisition of data: ZZ, RL, YW, YW YS, and CK. Analysis and interpretation of data: ZZ, RL, YS and RJ. Writing of the manuscript: ZZ. Study supervision: XW and JL.

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Correspondence to Jingtao Luo.

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Zhang, Z., Liu, R., Wang, Y. et al. Phosphorylation of MICAL2 by ARG promotes head and neck cancer tumorigenesis by regulating skeletal rearrangement. Oncogene 41, 334–346 (2022). https://doi.org/10.1038/s41388-021-02101-z

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