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Fbw7 promotes ubiquitin-dependent degradation of c-Myb: involvement of GSK3-mediated phosphorylation of Thr-572 in mouse c-Myb

Oncogene volume 28, pages 2393–2405 (2009)Cite this article

Abstract

Expression of oncoprotein c-Myb oscillates during hematopoiesis and hematological malignancies. Its quantity is not only regulated through transcriptional control but also through the ubiquitin–proteasome pathway, accompanied by phosphorylation, although the mechanisms are poorly understood. In this report, we tried to identify an E3 ubiquitin ligase, which targets c-Myb for ubiquitin-dependent degradation. We found that an F-box protein, Fbw7, interacted with c-Myb, which is mutated in numerous cancers. Fbw7 facilitated ubiquitylation and degradation of c-Myb in intact cells. Moreover, depletion of Fbw7 by RNA interference delayed turnover and increased the abundance of c-Myb in myeloid leukemia cells concomitantly, and suppressed the transcriptional level of γ-globin, which receives transcriptional repression from c-Myb. In addition, we analysed sites required for both ubiquitylation and degradation of c-Myb. We found that Thr-572 is critical for Fbw7-mediated ubiquitylation in mouse c-Myb using site-directed mutagenesis. Fbw7 recognized the phosphorylation of Thr-572, which was mediated by glycogen synthase kinase 3 (GSK3). In consequence, the c-Myb protein was markedly stabilized by the substitution of Thr-572 to Ala. These observations suggest that SCFFbw7 ubiquitin ligase regulates phosphorylation-dependent degradation of c-Myb protein.

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Acknowledgements

We thank Yojiro Kotake, Keiichi I Nakayama, Masaki Matsumoto, Akinobu Matsumoto and Issay Kitabayashi for providing plasmids and useful discussions, and Sayuri Suzuki, Tomomi Abe, Ning Liu, Harumi Shiratori, Daisuke Hiraoka, Daizo Ueno, Yasue Kirita, Tatsuya Kobayashi, Konomi Mizuguchi, Erina Nihashi, Syuhei Iizuka, Daisuke Ichikawa and Naohiro Takamoto for their technical assistance. This study was supported in part by grants from the Ministry of Education, Science, Sports, Culture and Technology of Japan (MK and KK), by a COE program of Hamamatsu University School of Medicine from the Ministry of Education, Science, Sports, Culture and Technology of Japan (MK).

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Author notes

  1. C Uchida & T Hattori

    Present address: Current address: Centre for Life Sciences, Oncology Research Institute, National University of Singapore, No. 02-0728 Medical Drive, Singapore 117456.,

Authors and Affiliations

  1. Department of Biochemistry 1, Hamamatsu University School of Medicine, Higashi-ku, Hamamatsu, Shizuoka, Japan,

    K Kitagawa, Y Hiramatsu, C Uchida, T Isobe, T Hattori, T Oda & M Kitagawa

  2. Second Department of Surgery, Hamamatsu University School of Medicine, Higashi-ku, Hamamatsu, Shizuoka, Japan,

    Y Hiramatsu

  3. Research Equipment Center, Hamamatsu University School of Medicine, Higashi-ku, Hamamatsu, Shizuoka, Japan,

    K Shibata

  4. Third Department of Internal Medicine, Hamamatsu University School of Medicine, Higashi-ku, Hamamatsu, Shizuoka, Japan,

    S Nakamura

  5. Department of Biochemistry, Graduate School of Biomedical Sciences, Hiroshima University, Minami-ku, Hiroshima, Japan,

    A Kikuchi

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  1. K Kitagawa
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Correspondence to M Kitagawa.

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Kitagawa, K., Hiramatsu, Y., Uchida, C. et al. Fbw7 promotes ubiquitin-dependent degradation of c-Myb: involvement of GSK3-mediated phosphorylation of Thr-572 in mouse c-Myb. Oncogene 28, 2393–2405 (2009). https://doi.org/10.1038/onc.2009.111

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