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Regulation of soluble Flt-1 (VEGFR-1) production by hnRNP D and protein arginine methylation

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Abstract

Soluble fms-like tyrosine kinase-1 (sFlt-1) functions as a potent inhibitor of angiogenesis by trapping vascular endothelial growth factor (VEGF). However, the precise regulatory mechanism of sFlt-1 production is unknown. Here, we report that vascular sFlt-1 production is regulated by heterogeneous nuclear ribonucleoprotein D (hnRNP D) and arginine methylation. We showed that hnRNP D bound to Flt-1 pre-mRNA and that hnRNP D overexpression decreased sFlt-1 mRNA in human microvascular endothelial cells (HMVECs). In contrast, the reduction of hnRNP D levels induced an increase in sFlt-1 production. Overexpression of an hnRNP D mutant in which the arginine residue of the known arginine methylation motif (arginine-glycine-glycine; RGG) was replaced with alanine did not reduce the level of soluble-form RNA produced from the Flt-1 minigene. Moreover, we demonstrated that overexpression of arginine methyltransferase decreased the soluble-form RNA level, whereas overexpression of arginine demethylase and addition of methyltransferase inhibitors increased sFlt-1 mRNA levels. These findings indicate that hnRNP D and arginine methylation play important roles in the regulation of Flt-1 mRNA alternative polyadenylation.

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Acknowledgments

This work was supported in part by a Grant-in-Aid for Young Scientists (B) (JSPS KAKENHI Grant Number 23792017), Strategic Research Foundation Grant-aided Project for Private Universities from The Ministry of Education, Culture, Sports, Science and Technology, Japan (S1201022), Grant for Specially Prompted Research from Kanazawa Medical University (SR2012-02), and Grant for Promoted Research from Kanazawa Medical University (S2013-11).

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Authors and Affiliations

  1. Department of Biochemistry, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, 920-0293, Japan

    Takayuki Ikeda, Yasuo Yoshitomi, Hidehito Saito, Yoshino Yoshitake & Hideto Yonekura

  2. Medical Research Institute, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, Japan

    Takeo Shimasaki, Yasuhito Ishigaki & Naohisa Tomosugi

  3. Department of Nephrology, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, Japan

    Hideki Yamaya

  4. Department of Cardiovascular Surgery, Kanazawa Medical University School of Medicine, 1-1 Daigaku, Uchinada, Kahoku-gun, Ishikawa, Japan

    Takashi Kobata

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  1. Takayuki Ikeda
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  2. Yasuo Yoshitomi
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  3. Hidehito Saito
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Correspondence to Hideto Yonekura.

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Ikeda, T., Yoshitomi, Y., Saito, H. et al. Regulation of soluble Flt-1 (VEGFR-1) production by hnRNP D and protein arginine methylation. Mol Cell Biochem 413, 155–164 (2016). https://doi.org/10.1007/s11010-015-2649-y

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  • DOI: https://doi.org/10.1007/s11010-015-2649-y

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