Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1

Lei Jiang, Daolin Tang, Kangkai Wang, Huali Zhang, Can Yuan, Dayue Duan, Xianzhong Xiao

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

A novel rat gene, Mipu1, encodes a 608 amino acid protein with an amino-terminal KRAB domain and 14 carboxyl-terminal C2H2 zinc finger motifs. Mipu1 is localized to the nucleus through its KRAB domain or the linker adjacent to its zinc finger region. Using the GST-Mipu1 bound to glutathione-Sepharose beads, a consensus putative DNA binding site (5′-TGTCTTATCGAA-3′) was extracted from a random oligonucleotide library. EMSA and target detection assay showed that the probe containing the putative site can bind to purified GST-Mipu1 fusion protein. The oligonucleotide containing the putative site was inserted into the pGL3-promotor vector to produce a reporter construct. The expression of reporter gene was repressed by overexpression of Mipu1 in a dose-dependent manner. Mutation analysis of the consensus sequence indicated that the repression mediated by Mipu1 is sequence-dependent. These results suggest that Mipu1 is a nuclear protein, which functions as a transcriptional repressor.

Original languageEnglish (US)
Pages (from-to)829-835
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume356
Issue number4
DOIs
StatePublished - May 18 2007
Externally publishedYes

Keywords

  • CH zinc finger
  • KRAB domain
  • Mipu1
  • Nuclear localization
  • Transcription factor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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