The zinc finger domain of Tzfp binds to the tbs motif located at the upstream flanking region of the Aie1 (aurora-C) kinase gene.
Our previous studies showed that Aie1 (aurora-C), is a novel testis kinase belonging to the aurora kinase family (). In this report, we describe a testis zinc finger protein (Tzfp) that binds to the upstream flanking sequence of the Aie1 gene. The mouse Tzfp gene, mapped to chromosome 7 B2-B3, encodes a 465-amino acid transcription factor containing a conserved N-terminal BTB/POZ domain and three C-terminal PLZF-like C(2)H(2) zinc fingers. The zinc finger domain of Tzfp binds to the TGTACAGTGT motif (Tzfp binding site, termed tbs) located at the upstream flanking sequence of the Aie1 gene by gel mobility shift, DNase I footprinting, and competition analyses. When the C-terminal zinc fingers of Tzfp were fused to the transactivation domain of VP16, the chimera activated transcription of a reporter construct containing multiple copies of the tbs. In contrast, the same chimera did not activate the reporter gene when an essential nucleotide fifth C was mutated to A at the tbs. Furthermore, we showed that the N-terminal BTB/POZ domain of TZFP has a repressor activity. Taken together, our results indicate that Tzfp recognizes a sequence-specific motif (tbs) and may play a role in the regulation of the genes carrying the tbs.
Tang
C J
CJ
Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, Republic of China.
Chuang
C K
CK
Hu
H M
HM
Tang
T K
TK
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D064108:Aurora Kinase C; D064127:Aurora Kinases; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D017930:Genes, Reporter; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D020033:Protein Isoforms; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D012097:Repressor Proteins; D017386:Sequence Homology, Amino Acid; D013737:Testis / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D016335:Zinc Fingers
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D012097:Repressor Proteins; C406863:Rog protein, mouse; D014157:Transcription Factors; C498646:ZBTB32 protein, human; C576927:AURKC protein, human; C576928:Aurkc protein, mouse; D064108:Aurora Kinase C; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases; D003851:Deoxyribonucleases