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Molecular and Cellular Biochemistry

, Volume 288, Issue 1–2, pp 151–157 | Cite as

The Spen Homolog Msx2-Interacting Nuclear Target Protein Interacts with the E2 Ubiquitin-Conjugating Enzyme UbcH8

  • Junfeng Li
  • Jishu Wang
  • Xi Yang
  • Junlin Li
  • Hongyan Qin
  • Xiao Dong
  • Yangting Zhu
  • Liang Liang
  • Yingmin Liang
  • Hua Han
Article

Abstract

The nuclear matrix protein Msx2-interacting nuclear target protein (MINT) is a transcription factor that regulates the expression of key transcriptional effectors in diverse signaling pathways. To further understand the function and mechanism of the MINT-mediated transcription regulation, the yeast two-hybrid system was employed to screen proteins that interact with the C-terminal fragment of MINT. From a cDNA library of human lymph nodes, a cDNA encoding the ubiquitin-conjugating enzyme UbcH8 was identified. Using different truncated versions of MINT, we show that the C-terminal Spen paralog and ortholog C-terminal domain (SPOC) domain, which has been demonstrated to mediate interactions between MINT and a panel of other molecules, might be responsible for interaction between MINT and UbcH8 in yeast, as confirmed by the β-galactosidase assay. The interaction between MINT and UbcH8 in mammalian cells was further proved by a series of biochemical assays including the mammalian two-hybrid assay, GST pull-down assay, and co-immunoprecipitation assay. Using a reporter system, we found that MINT-mediated transcription suppression was sensitive to MG132, an inhibitor of the proteosome system. These results suggest a novel mechanism of MINT-mediated transcription regulation, and might be helpful for understanding functions of MINT.

Key words

MINT transcription UbcH8 SPOC domain ubiquitin 

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Junfeng Li
    • 1
  • Jishu Wang
    • 1
  • Xi Yang
    • 1
  • Junlin Li
    • 1
  • Hongyan Qin
    • 1
  • Xiao Dong
    • 1
  • Yangting Zhu
    • 1
  • Liang Liang
    • 1
  • Yingmin Liang
    • 1
  • Hua Han
    • 1
  1. 1.State Key Laboratory of Cancer Biology, Department of Medical Genetics and Developmental BiologyFourth Military Medical UniversityXianP. R. China

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