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 journalArticle

17 Citations (Scopus)

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

Fingerprint

Functional analysis
Zinc Fingers
Oligonucleotides
Zinc
Genes
Consensus Sequence
Nuclear Proteins
Target tracking
Reporter Genes
Sepharose
Glutathione
Rats
Assays
Proteins
Fusion reactions
Binding Sites
Amino Acids
Mutation
DNA
CYS2-HIS2 Zinc Fingers

Keywords

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

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1. / Jiang, Lei; Tang, Daolin; Wang, Kangkai; Zhang, Huali; Yuan, Can; Duan, Dayue; Xiao, Xianzhong.

In: Biochemical and Biophysical Research Communications, Vol. 356, No. 4, 18.05.2007, p. 829-835.

Research output: Contribution to journalArticle

Jiang, Lei ; Tang, Daolin ; Wang, Kangkai ; Zhang, Huali ; Yuan, Can ; Duan, Dayue ; Xiao, Xianzhong. / Functional analysis of a novel KRAB/C2H2 zinc finger protein Mipu1. In: Biochemical and Biophysical Research Communications. 2007 ; Vol. 356, No. 4. pp. 829-835.
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