MBP-1 is efficiently encoded by an alternative transcript of the ENO1 gene but post-translationally regulated by proteasome-dependent protein turnover

FEBS J. 2010 Oct;277(20):4308-21. doi: 10.1111/j.1742-4658.2010.07819.x. Epub 2010 Sep 16.

Abstract

The c-myc promoter-binding protein-1 (MBP-1) is a transcriptional suppressor of tumorigenesis and thought to be the product of alternative translation initiation of the α-enolase (ENO1) transcript. In the present study, we cloned a 2552-bp novel cDNA with a putative coding sequence of MBP-1 and functionally examined its ability to encode the MBP-1 protein. Similarly to ENO1, the obtained MBP-1 was widely and differentially expressed in a variety of normal tissues and cancer cells. Experiments using MBP-1 promoter-driven luciferase reporter assays, biochemical cell fractionation followed by RT-PCR detection of the cytoplasmic mRNA, and transcription/translation-coupled reactions, consistently demonstrated that this novel transcript was alternatively transcribed from intron III of the ENO1 gene and was feasible for MBP-1 production. Hypoxia treatments significantly increased the transcriptional activation of the MBP-1 gene. Blocking the proteasomal degradation by MG132 stabilized the MBP-1 protein in cells. Compared with the translation efficiency for production of the MBP-1 protein, the MBP-1 transcript was 17.8 times more efficient than the ENO1 transcript. Thus, we suggest that this newly discovered transcript is a genuine template for the protein synthesis of MBP-1 in cells, and optimal expression of this gene in tumors may lead to effective clinical therapies for cancers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers, Tumor / genetics*
  • Bronchi / cytology
  • Cells, Cultured
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism*
  • Epithelial Cells / metabolism*
  • Gene Expression Regulation*
  • Humans
  • Hypoxia
  • Introns
  • Phosphopyruvate Hydratase / genetics*
  • Proteasome Endopeptidase Complex / metabolism*
  • Proteins / metabolism
  • RNA, Messenger
  • Tumor Suppressor Proteins / genetics*

Substances

  • Biomarkers, Tumor
  • DENND4A protein, human
  • DNA-Binding Proteins
  • Proteins
  • RNA, Messenger
  • Tumor Suppressor Proteins
  • Proteasome Endopeptidase Complex
  • ENO1 protein, human
  • Phosphopyruvate Hydratase