cDNA cloning and transcriptional properties of a novel GC box-binding protein, BTEB2.
We have cloned a cDNA for a novel GC box-binding protein designated BTEB2 from a human placenta cDNA library using rat BTEB cDNA (Imataka et al. (1992). EMBO J. 11,3663-3671. as a hybridization probe. BTEB2 consists of 219 amino acids and contains three contiguous zinc finger motifs at its C-terminus. The zinc finger domains showed 59% and 64% sequence similarity to those of Sp1 and BTEB, respectively. Adjacent to the N-terminal of the zinc finger motifs, a short sequence rich in basic amino acids is conserved between BTEB2 and Sp1. Furthermore, This basic sequence concurs with the N-terminal half of the consensus sequence for basic domains of the proteins containing both helix-loop-helix and leucine zipper motifs. The other region of BTEB2 is notably rich in proline, serine, threonine, and alanine residues. BTEB2 expressed in Escherichia coli showed DNA-binding activity whose specificity was closely similar to that of Sp1. Cotransfection experiments using Hepa-1 cells (a mouse hepatoma cell line) with a BTEB2 expression plasmid and GC box-containing reporter plasmids revealed that BTEB2 apparently activated the expression of the CAT activity. Moreover, when BTEB2 was fused to GAL4 DNA-binding domain, the chimeric protein could enhance the transcription through promoters containing GAL4-binding sites. Analysis of the BTEB2 mRNA by RNA blot analysis demonstrated that the mRNA was expressed specifically in testis and placenta with different sizes, 20S and 28S, respectively, among various organs examined.
Sogawa
K
K
Department of Chemistry, Faculty of Science, Tohoku University, Sendai, Japan.
Imataka
H
H
Yamasaki
Y
Y
Kusume
H
H
Abe
H
H
Fujii-Kuriyama
Y
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics*; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D008969:Molecular Sequence Data; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000235:genetics*; D014157:Transcription Factors; D014158:Transcription, Genetic; D016335:Zinc Fingers / Q000235:genetics
D004268:DNA-Binding Proteins; C492214:KLF5 protein, human; C489676:KLF9 protein, human; C118391:Klf5 protein, mouse; C492215:Klf5 protein, rat; C489675:Klf9 protein, mouse; D051741:Kruppel-Like Transcription Factors; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C077017:Klf9 protein, rat; D015500:Chloramphenicol O-Acetyltransferase