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Control of cell migration by the novel protein phosphatase-2A interacting protein inka2

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Abstract

Cell migration is essential for many physiological and pathological processes, including embryonic development, wound healing, immune response and cancer metastasis. Inka2 transcripts are observed in migrating cells during embryonic development, suggesting the involvement of inka2 in cell migration. However, its precise role remains unclear. Here, we found that inka2 controlled focal adhesion dynamics and cell migration, likely by regulating protein phosphatase-2A (PP2A) function. A scratch assay revealed that inka2 shRNA-transfected NIH3T3 cells showed rapid wound closure, indicating an inhibitory effect by inka2 on cell migration. Live-cell imaging of NIH3T3 cells expressing EGFP-paxillin using total internal reflection fluorescence microscopy revealed that inka2 knockdown increased the turnover rate of focal adhesions. Given that PP2A, which consists of catalytic (C), regulatory (B) and scaffolding (A) subunits, is known to regulate focal adhesions, we examined the inka2-PP2A interaction. Immunoprecipitation revealed an association between inka2 and the PP2A C subunit. Binding of Inka2 to the C subunit prevented the association between the A and C subunits, suggesting that inka2 can inhibit PP2A function. Furthermore, both inka2 expression and PP2A inhibition decreased focal adhesion kinase-paxillin interaction, resulting in reduced formation of focal adhesions. We assessed the effect of pharmacological PP2A inhibition on the inka2 knockdown-induced increase in cell migration speed and found that treatment with a PP2A inhibitor negated the accelerated migration of inka2 knockdown cells. These results suggest that inka2 knockdown exerts its effects through PP2A-dependent regulation of focal adhesions. Our findings contribute to a better understanding of the molecular mechanisms underlying cell migration.

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Acknowledgments

We are grateful to Prof. W. Hahn for providing plasmids, DONR223_PPP2R1A_WT and pBABE zeo PPP2CA Wt, obtained from Addgene. We thank Dr. T. Hida, Ms. R. Shigematsu, and Ms. A. Tanaka for their technical assistance.

We would like to thank Editage (www.editage.jp) for English language editing.

Funding

This work was supported by JSPS KAKENHI grant numbers 16 K14583, 18 K06491 (to H.A.) and 26430042 (to S.S.) and by Waseda University Grants for Special Research Projects 2018 K-352 (to H.A.), 2015 K-249 and 2017 K-301 (to S.S.).

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Authors and Affiliations

  1. Laboratory for Molecular Neurobiology, Faculty of Human Sciences, Waseda University, Tokorozawa, Saitama, 359-1192, Japan

    Hiroki Akiyama, Yumi Iwasaki, Seiya Yamada & Shin-ichi Sakakibara

  2. Laboratory for Neural Cell Dynamics, RIKEN Center for Brain Science, Wako, Saitama, 351-0198, Japan

    Hiroki Akiyama & Hiroyuki Kamiguchi

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  1. Hiroki Akiyama
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  2. Yumi Iwasaki
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  3. Seiya Yamada
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  4. Hiroyuki Kamiguchi
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  5. Shin-ichi Sakakibara
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Correspondence to Hiroki Akiyama or Shin-ichi Sakakibara.

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All experiments in this study were approved by the Committee on the Ethics of Animal Experiments of Waseda University.

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Akiyama, H., Iwasaki, Y., Yamada, S. et al. Control of cell migration by the novel protein phosphatase-2A interacting protein inka2. Cell Tissue Res 380, 527–537 (2020). https://doi.org/10.1007/s00441-020-03169-x

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