Induction of amphiregulin by p53 promotes apoptosis via control of microRNA biogenesis in response to DNA damage

Edited by Moshe Oren, Weizmann Institute of Science, Rehovot, Israel, and accepted by the Editorial Board December 11, 2013 (received for review July 20, 2013)
December 30, 2013
111 (2) 717-722

Significance

The tumor suppressive function of p53 is tightly regulated by its posttranslational modifications. Although Ser46 phosphorylation is a critical modification for apoptosis induction, a molecular mechanism by which Ser46-phosphorylated p53 induces apoptosis remains unclear. Here, we clarify that amphiregulin (AREG) is specifically induced in a Ser46 phosphorylation-specific manner. Notably, AREG colocalizes with DEAD-box RNA helicase p68 (DDX5) in the nucleus and regulates tumor suppressive microRNA biogenesis in response to DNA damage. These findings support a model in which Ser46-phosphorylated p53 orchestrates tumor suppressive microRNA expression in the apoptotic response to DNA damage.

Abstract

Upon DNA damage, tumor suppressor p53 determines cell fate by repairing DNA lesions to survive or by inducing apoptosis to eliminate damaged cells. The decision is based on its posttranslational modifications. Especially, p53 phosphorylation at Ser46 exerts apoptotic cell death. However, little is known about the precise mechanism of p53 phosphorylation on the induction of apoptosis. Here, we show that amphiregulin (AREG) is identified for a direct target of Ser46 phosphorylation via the comprehensive expression analyses. Ser46-phosphorylated p53 selectively binds to the promoter region of AREG gene, indicating that the p53 modification changes target genes by altering its binding affinity to the promoter. Although AREG belongs to a family of the epidermal growth factor, it also emerges in the nucleus under DNA damage. To clarify nuclear function of AREG, we analyze AREG-binding proteins by mass spectrometry. AREG interacts with DEAD-box RNA helicase p68 (DDX5). Intriguingly, AREG regulates precursor microRNA processing (i.e., miR-15a) with DDX5 to reduce the expression of antiapoptotic protein Bcl-2. These findings collectively support a mechanism in which the induction of AREG by Ser46-phosphorylated p53 is required for the microRNA biogenesis in the apoptotic response to DNA damage.

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Data Availability

Data deposition: The complete expression datasets have been deposited in the Array Express database, www.ebi.ac.uk/arrayexpress (accession no. E-MEXP-2556).

Acknowledgments

We thank Ayako Igarashi for technical assistance. This work was supported by grants from the Ministry of Education, Science and Culture of Japan, the Jikei University Graduate Research Fund, the NOVARTIS Foundation for the Promotion of Science, Suzuken Memorial Foundation, Uehara Memorial Foundation, the Astellas Foundation for Research on Medical Resources, Takeda Science Foundation, the Mochida Memorial Foundation for Medical and Pharmaceutical Research, the Sumitomo Foundation, the Naito Foundation, Japan Foundation for Applied Enzymology, Project Mirai Cancer Research Grants, the Yasuda Memorial Foundation, the Japan Society for the Promotion of Science, and the Nakajima Foundation.

Supporting Information

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Supporting Information

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 111 | No. 2
January 14, 2014
PubMed: 24379358

Classifications

Data Availability

Data deposition: The complete expression datasets have been deposited in the Array Express database, www.ebi.ac.uk/arrayexpress (accession no. E-MEXP-2556).

Submission history

Published online: December 30, 2013
Published in issue: January 14, 2014

Keywords

  1. microarray
  2. Drosha
  3. miRNA processing

Acknowledgments

We thank Ayako Igarashi for technical assistance. This work was supported by grants from the Ministry of Education, Science and Culture of Japan, the Jikei University Graduate Research Fund, the NOVARTIS Foundation for the Promotion of Science, Suzuken Memorial Foundation, Uehara Memorial Foundation, the Astellas Foundation for Research on Medical Resources, Takeda Science Foundation, the Mochida Memorial Foundation for Medical and Pharmaceutical Research, the Sumitomo Foundation, the Naito Foundation, Japan Foundation for Applied Enzymology, Project Mirai Cancer Research Grants, the Yasuda Memorial Foundation, the Japan Society for the Promotion of Science, and the Nakajima Foundation.

Notes

This article is a PNAS Direct Submission. M.O. is a guest editor invited by the Editorial Board.

Authors

Affiliations

Naoe Taira
Department of Biochemistry, The Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo 105-8461, Japan;
Tomoko Yamaguchi
Department of Biochemistry, The Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo 105-8461, Japan;
Junko Kimura
Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan;
Zheng-Guang Lu
Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8510, Japan;
Shinji Fukuda
Department of Chemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime 791-0295, Japan; and
Shigeki Higashiyama
Department of Chemistry and Molecular Genetics, Ehime University Graduate School of Medicine, Shitsukawa, Toon, Ehime 791-0295, Japan; and
Masaya Ono
Chemotherapy Division and Cancer Proteomics Project, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan
Kiyotsugu Yoshida1 [email protected]
Department of Biochemistry, The Jikei University School of Medicine, 3-25-8 Nishi-shinbashi, Minato-ku, Tokyo 105-8461, Japan;

Notes

1
To whom correspondence should be addressed. E-mail: [email protected].
Author contributions: N.T. and K.Y. designed research; N.T., T.Y., J.K., Z.-G.L., and M.O. performed research; S.F. and S.H. contributed new reagents/analytic tools; N.T. and K.Y. analyzed data; and N.T. and K.Y. wrote the paper.

Competing Interests

The authors declare no conflict of interest.

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    Induction of amphiregulin by p53 promotes apoptosis via control of microRNA biogenesis in response to DNA damage
    Proceedings of the National Academy of Sciences
    • Vol. 111
    • No. 2
    • pp. 565-876

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