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Digestive Diseases and Sciences

, Volume 57, Issue 9, pp 2347–2354 | Cite as

Identification of a Novel Binding Protein of FAT10: Eukaryotic Translation Elongation Factor 1A1

  • Xin Yu
  • Xiuxia Liu
  • Tiande Liu
  • Kui Hong
  • Jun Lei
  • Rongfa Yuan
  • Jianghua Shao
Original Article

Abstract

Background

FAT10 is known to execute its functions mainly through conjugation to different substrates, and these known functions include cytokine responses, apoptosis, mitosis, and tumorigenesis. Nonetheless, the known binding proteins of FAT10 cannot explain all its known functions. As such, the aim of this study was to identify unidentified conjugation proteins of FAT10.

Methods

The yeast two-hybrid system was employed in this study. FAT10 was used as the bait protein for screening of a cDNA library from a human hepatocellular carcinoma cell line, Hep3B. Protein interactions were confirmed based on localization studies and co-immunoprecipitation assays. The expression of mRNA and protein was determined using real-time polymerase chain reaction and western blot analyses, respectively.

Results

In this study, we identified eukaryotic elongation factor 1A1 (eEF1A1) as a FAT10-specific binding protein. The binding between FAT10 and eEF1A1 was confirmed both in vivo and in vitro. We also found that, when the expression of FAT10 was reduced by siRNA knockdown, this resulted in downregulation of eEF1A1 expression at both the mRNA and protein levels in human hepatocellular carcinoma cells.

Conclusions

We propose a model in which eEF1A1 serves as a substrate of FAT10 to accomplish, in part, its functions in regulating the biological behavior of tumor cells. Since both eEF1A1 and FAT10 are important for tumorigenesis and development, comprehending the mechanisms of this interaction can provide clues for identification of novel strategic targets for drug screening and molecular typing, and possibly in the development of new effective therapeutic strategies against hepatocellular carcinoma.

Keywords

Human HLA-F adjacent transcript 10 (FAT10) Eukaryotic translation elongation factor 1A1 (eEF1A1) Yeast two-hybrid system siRNA 

Notes

Acknowledgments

This work was supported by grants from the Key Project of Chinese Ministry of Education (208070) and the National Natural Science Foundation of China (81060196).

Conflict of interest

None.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Xin Yu
    • 1
  • Xiuxia Liu
    • 1
    • 2
  • Tiande Liu
    • 1
  • Kui Hong
    • 2
  • Jun Lei
    • 1
  • Rongfa Yuan
    • 1
  • Jianghua Shao
    • 1
    • 2
  1. 1.The Department of Hepatobiliary SurgeryThe Second Affiliated Hospital of Nanchang UniversityNanchangChina
  2. 2.The Key Laboratory of Molecular MedicineThe Second Affiliated Hospital of Nanchang UniversityNanchangChina

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