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CRL2KLHDC3 and CRL1Fbxw7 cooperatively mediate c-Myc degradation

Oncogene (2024)Cite this article

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Abstract

c-Myc is a proto-oncoprotein that regulates various cellular processes and whose abnormal expression leads to tumorigenesis. c-Myc protein stability has been shown to be predominantly controlled by the ubiquitin ligase (E3) CRL1Fbxw7 in a manner dependent on glycogen synthase kinase 3 (GSK3)–mediated phosphorylation. Here we show that, in some types of cancer cells, c-Myc degradation is largely insensitive to the GSK3 inhibitor (GSK3i) CHIR99021, suggesting the existence of an E3 other than CRL1Fbxw7 for c-Myc degradation. Mass spectrometry identified CRL2KLHDC3 as such an E3. In GSK3i-insensitive cancer cells, combined depletion of Fbxw7 and KLHDC3 resulted in marked stabilization of c-Myc, suggestive of a cooperative action of Fbxw7 and KLHDC3. Furthermore, transplantation of such cells deficient in both Fbxw7 and KLHDC3 into immunodeficient mice gave rise to larger tumors compared with those formed by cells lacking only Fbxw7. GSK3i-insensitive pancreatic cancer cells expressed lower levels of SHISA2, a negative regulator of the Wnt signaling pathway, than did GSK3i-sensitive cells. KLHDC3 mRNA abundance was associated with prognosis in pancreatic cancer patients with a low level of SHISA2 gene expression. These results suggest that KLHDC3 cooperates with Fbxw7 to promote c-Myc degradation in a subset of cancer cells with low GSK3 activity.

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Fig. 1: Some cancer cell lines are less dependent on Fbxw7 for c-Myc degradation.
Fig. 2: Identification of CRL2KLHDC3 as a candidate E3 for c-Myc.
Fig. 3: KLHDC3 mediates the interaction between CUL2 and c-Myc through binding to Myc box 2.
Fig. 4: CRL2KLHDC3 mediates c-Myc degradation.
Fig. 5: Translocation of KLHDC3 to the nucleus in a manner dependent on c-Myc binding.
Fig. 6: Loss of KLHDC3 accelerates tumor formation by Fbxw7 KO cells.
Fig. 7: GSK3 activity determines the dependence of c-Myc degradation on KLHDC3.
Fig. 8: Two E3s cooperatively target c-Myc for degradation.

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Data availability

All proteome data are available in ProteomeXchange and jPOSTrepo under the accession numbers PXD047181 and JPST002393, respectively.

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Acknowledgements

We thank members of the Department of Molecular and Cellular Biology as well as the Research Promotion Unit, Medical Institute of Bioregulation, Kyushu University, for technical assistance, as well as Y. Okabe, A. Matsukuma, K. Tsunematsu, and A. Ohta for help with preparation of the manuscript. Computations were performed in part on the NIG supercomputer at ROIS National Institute of Genetics. Analysis of RNA-sequencing data was performed with the pipeline provided by Rhelixa Inc. This research was supported in part by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan to KIN (JP23H00378) as well as from the Japan Agency for Medical Research and Development (AMED) to KIN (JP23ck0106802 and JP23ama221119) and from the FOREST Program of the Japan Science and Technology Agency (JST) to KY (JPMJFR216G).

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Authors and Affiliations

  1. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, 812-8582, Japan

    Saori Motomura, Kanae Yumimoto & Keiichi I. Nakayama

  2. Laboratory of Proteome Research, National Institutes of Biomedical Innovation, Health, and Nutrition, Ibaraki, Osaka, 567-0085, Japan

    Takeshi Tomonaga

  3. Anticancer Strategies Laboratory, TMDU Advanced Research Institute, Tokyo Medical and Dental University, Tokyo, 113-8510, Japan

    Keiichi I. Nakayama

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  1. Saori Motomura
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SM designed and performed experiments. KY conceived and designed the project. TT provided cell lines. KIN supervised the project. SM, KY, and KIN wrote the manuscript.

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Correspondence to Kanae Yumimoto or Keiichi I. Nakayama.

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Motomura, S., Yumimoto, K., Tomonaga, T. et al. CRL2KLHDC3 and CRL1Fbxw7 cooperatively mediate c-Myc degradation. Oncogene (2024). https://doi.org/10.1038/s41388-024-03048-7

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