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Identification of UAP1L1 as a critical factor for protein O-GlcNAcylation and cell proliferation in human hepatoma cells

Oncogene (2018) | Download Citation

Abstract

Aged hepatocyte-specific-Mcl-1 knockout (MKO-hep) mice are prone to develop liver tumors mimicking human hepatocellular carcinoma (HCC). Here we reported that a protein named UDP-N-acetylglucosamine pyrophosphorylase-1-like-1 (Uap1l1) is upregulated in the liver of young MKO-hep mice without any macroscopically detectable tumor nodules and is prominently expressed in the hepatic tumors developed in the aged MKO-hep mice. Intriguingly, human UAP1L1 is also significantly upregulated in a distinct subset of HCC tissues and patients with upregulated expression of UAP1L1 appeared to have poor prognosis. Overexpression of UAP1L1 significantly promoted, whereas UAP1L1 knockdown markedly reduced the proliferation of human hepatoma cells both in vitro and in vivo. UAP1L1 shows ~59% sequence identity to UDP-N-acetylglucosamine pyrophosphorylase-1 (UAP1), which is directly involved in the synthesis of the sugar donor (UDP-GlcNac) for N-acetylglucosamine modification (O-GlcNAcylation) of proteins. However, unlike UAP1, UAP1L1 harbors very limited UDP-GlcNAc synthesis activity. Moreover, although both UAP1 and UAP1L1 are required for O-GlcNAc transferase (OGT)-mediated protein O-GlcNAcylation, they appear to function distinctly from each other. UAP1L1 directly interacts with OGT, but does not seem to be an OGT substrate. In addition, UAP1L1 alone is not sufficient to activate OGT activity in vitro, suggesting that UAP1L1 may function together with other proteins to modulate OGT activity in vivo. Lastly, UAP1L1 knockdown attenuated c-MYC O-GlcNAcylation and protein stability, and overexpression of c-MYC significantly rescued the proliferation defect of UAP1L1 knockdown HepG2 cells, suggesting that c-MYC is one downstream target of UAP1L1 that contributes to UAP1L1-mediated cell proliferation, at least in HepG2 cells.

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Acknowledgements

We greatly appreciate the gift of mammalian OGT expression vectors from Dr Xiaochun Yu (Beckman Research Institute), the bacterial OGT expression vector from Dr Hsiu-Ming Shih (Academia Sinica), and various hepatoma cell lines from Dr Yuh-Shan Jou (Academia Sinica). We also thank Yi-Ning Chen at the Bioinformatics core at IMB for I-TASSER modeling and the Metabolomics Facilities of the Scientific Instrument Center at Academia Sinica for the analysis of the cellular levels of UDP-GlcNAc. This study was supported in part by an intramural fund from Academia Sinica and by grants (99-2320-B-001-009-MY3; 102-2320-B-001 -024 -MY3) from the National Science Council of Taiwan to H-FY-Y; funding to HL was provided by Chang Gung Memorial Hospital (CMRPG3D1512) and the Ministry of Education of Taiwan (EMRPD1E1581).

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Author notes

  1. These authors contributed equally: Ching-Yu Lai, Hsuan Liu, Kai Xuan Tin

Affiliations

  1. Institute of Molecular Biology, Academia Sinica, Taipei, Taiwan

    • Ching-Yu Lai
    • , Kai Xuan Tin
    • , Kun-Hai Yeh
    • , Hubert W. Peng
    • , Huan-Da Chen
    • , Jun-Yu He
    • , Shih-Yen Weng
    •  & Hsin-Fang Yang-Yen
  2. Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan

    • Ching-Yu Lai
    • , Mi-Hua Tao
    • , Jeffrey Jong-Young Yen
    •  & Hsin-Fang Yang-Yen
  3. Molecular Medicine Research Center, Chang Gung University, Tao-Yuan, Taiwan

    • Hsuan Liu
    •  & Yi Huang
  4. Department of Molecular and Cellular Biology, Chang Gung University, Tao-Yuan, Taiwan

    • Hsuan Liu
  5. ACT Genomics Co., LTD, Taipei, Taiwan

    • Yi Huang
  6. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    • Yun-Jung Chiang
    • , Chun-Shan Liu
    • , Mi-Hua Tao
    •  & Jeffrey Jong-Young Yen
  7. Division of Gastroenterology, Department of Internal Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan

    • Shih-Yen Weng

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Correspondence to Hsin-Fang Yang-Yen.

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DOI

https://doi.org/10.1038/s41388-018-0442-6