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E2-EPF UCP targets pVHL for degradation and associates with tumor growth and metastasis

Nature Medicine volume 12, pages 809–816 (2006)Cite this article

Abstract

The von Hippel-Lindau tumor suppressor, pVHL, forms part of an E3 ubiquitin ligase complex that targets specific substrates for degradation, including hypoxia-inducible factor-1α (HIF-1α), which is involved in tumor progression and angiogenesis. It remains unclear, however, how pVHL is destabilized. Here we show that E2-EPF ubiquitin carrier protein (UCP) associates with and targets pVHL for ubiquitin-mediated proteolysis in cells, thereby stabilizing HIF-1α. UCP is detected coincidently with HIF-1α in human primary liver, colon and breast tumors, and metastatic cholangiocarcinoma and colon cancer cells. UCP level correlates inversely with pVHL level in most tumor cell lines. In vitro and in vivo, forced expression of UCP boosts tumor-cell proliferation, invasion and metastasis through effects on the pVHL-HIF pathway. Our results suggest that UCP helps stabilize HIF-1α and may be a new molecular target for therapeutic intervention in human cancers.

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Figure 1: UCP interacts with pVHL, but not the pVHL E3 ubiquitin ligase complex.
Figure 2: UCP targets pVHL for ubiquitin-mediated proteolysis, resulting in an accumulation of HIF-1α.
Figure 3: UCP stabilizes HIF-1α in the mouse liver and is coincidently detected with HIF-1α in human tumor cells.
Figure 4: UCP promotes tumor cell proliferation and invasion in culture.
Figure 5: UCP enhances tumor growth and metastasis in mice, which are inhibited by treatment with Ad.UCP-siRNA.
Figure 6: The effects of UCP on tumor-cell proliferation and invasion are mediated by the pVHL-HIF pathway.

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Acknowledgements

We are grateful to the members of the Center for Functional Analysis of Human Genome and 21C Frontier Human Gene Bank, Republic of Korea; Y.I. Yeom and N.S. Kim for providing full-length cDNAs; J.H. Lee for providing invasion and metastasis assays and C8161 cell line; D.G. Kim, I.P. Choi, and Y.S. Kim for providing tumor cell lines; C.K. Lee and D.Y. Yu for the use of animal facility. We also thank the reviewers for helpful remarks that improved this manuscript. This work was supported by a grant of 21C Frontier Functional Genome Project from the Ministry of Science and Technology, Republic of Korea.

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Authors and Affiliations

  1. Gene Therapy Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yusong-gu, 305-333, Daejeon, Republic of Korea

    Cho-Rok Jung, Kyung-Sun Hwang, Jinsang Yoo, Won-Kyung Cho & Dong-Soo Im

  2. Department of Pathology, Chung-Nam National University School of Medicine, Chung-gu, 301-721, Daejeon, Republic of Korea

    Jin-Man Kim

  3. Department of Pathology, Seoul National University College of Medicine, Jongro-gu, 110-799, Seoul, Republic of Korea

    Woo Ho Kim

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Supplementary information

Supplementary Fig. 1

F-UCP expressed in UCP-negative cells co-sediments with pVHL. (PDF 98 kb)

Supplementary Fig. 2

UCP targets pVHL in cells under hypoxic conditions, thereby accumulating active HIF-1α. (PDF 89 kb)

Supplementary Fig. 3

Recombinant GST-UCP, but not GST-UCPm is catalytically active. (PDF 90 kb)

Supplementary Fig. 4

UCP ubiquitinates pVHL in vitro (PDF 119 kb)

Supplementary Fig. 5

UCP specifically targets pVHL for degradation. (PDF 125 kb)

Supplementary Fig. 6

UCP targets pVHL for degradation in Ck-K1 cells, thereby stabilizing HIF-1α. (PDF 85 kb)

Supplementary Fig. 7

The specificity for UCP-siRNA. (PDF 114 kb)

Supplementary Fig. 8

Detection of Ad.F-UCP genome and F-UCP transcript in tumors excised from mice. (PDF 111 kb)

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Jung, CR., Hwang, KS., Yoo, J. et al. E2-EPF UCP targets pVHL for degradation and associates with tumor growth and metastasis. Nat Med 12, 809–816 (2006). https://doi.org/10.1038/nm1440

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