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Identification of the ubiquitin–protein ligase that recognizes oxidized IRP2

Nature Cell Biology volume 5, pages 336–340 (2003)Cite this article

Abstract

The ubiquitin system is involved in several basic cellular functions1,2,3. Ubiquitination is carried out by a cascade of three reactions catalysed by the E1, E2 and E3 enzymes. Among these, the E3 ubiquitin–protein ligases have a pivotal role in determining the specificity of the system by recognizing the target substrates through defined targeting motifs1,2,3. Although RING finger proteins constitute an important family of E3 ligases4, only a few post-transcriptional modifications, including phosphorylation1, proline hydroxylation5,6 and glycosylation7, are known to function as recognition signals for E3. Iron regulatory protein 2 (IRP2), a modulator of iron metabolism, is regulated by iron-induced ubiquitination and degradation8. Here we show that the RING finger protein HOIL-1 functions as an E3 ligase for oxidized IRP2, suggesting that oxidation is a specific recognition signal for ubiquitination. The oxidation of IRP2 is generated by haem, which binds to IRP2 in iron-rich cells, and by oxygen, indicating that the iron sensing of IRP2 depends on the synthesis and availability of haem.

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Figure 1: HOIL-1 binds IRP2 in the presence of oxygen.
Figure 2: HOIL-1 acts as a ubiquitin–protein ligase for oxidized IRP2 in vitro.
Figure 3: HOIL-1 functions as an IRP2-E3 in cells.
Figure 4: HOIL-1 targeting of IRP2 for degradation requires haem and oxygen.

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Acknowledgements

We thank K. Tanaka for critically reading the manuscript; A. Dancis for discussions; and R. Klausner for reagents. This work was partly supported by grants from Ministry of Education, Culture, Sports, Science and Technology of Japan to K.I.

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Author notes

  1. Koji Yamanaka and Haruto Ishikawa: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Molecular & System Biology, Graduate School of Biostudies, Kyoto University, Sakyo-Ku, Kyoto, 606-8501, Japan

    Koji Yamanaka, Nagahiro Minato & Kazuhiro Iwai

  2. Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Sakyo-Ku, Kyoto, 606-8501, Japan

    Haruto Ishikawa, Isao Morishima & Koichiro Ishimori

  3. Department of Molecular Cell Biology, Graduate School of Medicine, Osaka City University, 1-4-3 Asahi-Machi, Abeno-Ku, Osaka, 545-8585, Japan

    Haruto Ishikawa, Yuzuru Megumi, Fuminori Tokunaga, Masato Kanie & Kazuhiro Iwai

  4. CREST, Japan Science and Technology Corporation (JST), Kawaguchi, 332-0012, Saitama, Japan

    Haruto Ishikawa, Fuminori Tokunaga, Koichiro Ishimori & Kazuhiro Iwai

  5. Cell Biology & Metabolism Branch, National Institute of Child Health and Human Development, Bethesda, 20892-5430, Maryland, USA

    Tracey A. Rouault

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  1. Koji Yamanaka
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Correspondence to Kazuhiro Iwai.

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Yamanaka, K., Ishikawa, H., Megumi, Y. et al. Identification of the ubiquitin–protein ligase that recognizes oxidized IRP2. Nat Cell Biol 5, 336–340 (2003). https://doi.org/10.1038/ncb952

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