Cloning and characterization of a novel human zinc finger gene, hKid3, from a C2H2-ZNF enriched human embryonic cDNA library.
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.
Gao
Li
L
Ministry of Education Key Laboratory of Virology, College of Life Science, Wuhan University, Wuhan, Hubei 430072, PR China. liwxlab@whu.edu.cn
Sun
Chong
C
Qiu
Hong-Ling
HL
Liu
Hui
H
Shao
Huan-Jie
HJ
Wang
Jun
J
Li
Wen-Xin
WX
D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D007668:Kidney / Q000196:embryology / Q000378:metabolism*; D051741:Kruppel-Like Transcription Factors; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000235:genetics*; D009928:Organ Specificity; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics*; D020539:Sequence Analysis, Protein / Q000379:methods; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014018:Tissue Distribution; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics
C486563:BCL11A protein, human; D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D051741:Kruppel-Like Transcription Factors; D009687:Nuclear Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; C485805:ZNF354C protein, human