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The mammalian gene ZNF268 is regulated by hUpf1

Chengang Zhu1, Zhouzhou Zhao, Mingxiong Guo

  • 1State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, P. R. China.

Biochemistry. Biokhimiia
|September 9, 2008
PubMed

Insights

Nonsense-mediated mRNA decay (NMD) regulates the mammalian ZNF268 gene promoter via the hUpf1 protein. This study reveals NMD

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • RNA Surveillance

Background:

  • Nonsense-mediated mRNA decay (NMD) degrades aberrant mRNAs with premature stop codons.
  • NMD is known to regulate gene expression transcriptionally in yeast.
  • Transcriptional regulation by NMD in higher eukaryotes, particularly at the promoter level, remains largely unexplored.

Purpose of the Study:

  • To investigate the potential role of the NMD pathway in regulating the ZNF268 gene promoter in mammalian cells.
  • To elucidate the specific NMD factors involved in ZNF268 promoter regulation.

Main Methods:

  • Transient transfection assays to assess ZNF268 promoter activity.
  • Depletion of endogenous NMD factors (hUpf1, hSmg6) using siRNA in HeLa cells.
  • Analysis of steady-state ZNF268 mRNA levels.
  • Site-directed mutagenesis of the ZNF268 promoter, focusing on the C/EBP site.

Main Results:

  • ZNF268 promoter activity was significantly influenced by hUpf1, but not hSmg6, in HeLa cells.
  • Depletion of endogenous hUpf1, but not hSmg6, affected ZNF268 mRNA levels, confirming the promoter activity findings.
  • Mutational analysis identified the C/EBP transcription factor binding site as crucial for hUpf1-mediated regulation of the ZNF268 promoter.

Conclusions:

  • The mammalian gene ZNF268 is transcriptionally regulated by the nonsense-mediated mRNA decay pathway.
  • The hUpf1 protein plays a key role in this regulation, acting through the ZNF268 promoter.
  • The C/EBP binding site is a critical element mediating hUpf1's regulatory effect on the ZNF268 promoter.

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