Cloning and characterization of a novel human zinc finger gene, hKid3, from a C2H2-ZNF enriched human embryonic cDNA library

Biochem Biophys Res Commun. 2004 Dec 24;325(4):1145-52. doi: 10.1016/j.bbrc.2004.10.158.

Abstract

To investigate the zinc finger genes involved in human embryonic development, we constructed a C(2)H(2)-ZNF enriched human embryonic cDNA library, from which a novel human gene named hKid3 was identified. The hKid3 cDNA encodes a 554 amino acid protein with an amino-terminal KRAB domain and 11 carboxyl-terminal C(2)H(2) zinc finger motifs. Northern blot analysis indicates that two hKid3 transcripts of 6 and 8.5kb express in human fetal brain and kidney. The 6kb transcript can also be detected in human adult brain, heart, and skeletal muscle while the 8.5kb transcript appears to be embryo-specific. GFP-fused hKid3 protein is localized to nuclei and the ZF domain is necessary and sufficient for nuclear localization. To explore the DNA-binding specificity of hKid3, an oligonucleotide library was selected by GST fusion protein of hKid3 ZF domain, and the consensus core sequence 5'-CCAC-3' was evaluated by competitive electrophoretic mobility shift assay. Moreover, The KRAB domain of hKid3 exhibits transcription repressor activity when tested in GAL4 fusion protein assay. These results indicate that hKid3 may function as a transcription repressor with regulated expression pattern during human development of brain and kidney.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Brain / embryology
  • Brain / metabolism*
  • Carrier Proteins / chemistry*
  • Carrier Proteins / genetics*
  • Cells, Cultured
  • Cloning, Molecular / methods*
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics*
  • Gene Library
  • Humans
  • Kidney / embryology
  • Kidney / metabolism*
  • Kruppel-Like Transcription Factors
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / genetics*
  • Organ Specificity
  • Repressor Proteins / chemistry*
  • Repressor Proteins / genetics*
  • Sequence Analysis, Protein / methods
  • Sequence Homology, Amino Acid
  • Species Specificity
  • Tissue Distribution
  • Transcription Factors / chemistry
  • Transcription Factors / genetics

Substances

  • BCL11A protein, human
  • Carrier Proteins
  • DNA-Binding Proteins
  • Kruppel-Like Transcription Factors
  • Nuclear Proteins
  • Repressor Proteins
  • Transcription Factors
  • ZNF354C protein, human