Candidate cis-elements for human renin gene expression in the promoter region.
The regulation of renin gene expression, the rate-limiting enzyme of the system, is thought to be fundamental to the total system. Previously, we mapped six putative cis-elements in the promoter region of the human renin gene with nuclear proteins from human chorionic cells and human renal cortex by DNase I protection assay (footprint A-F). Each footprint contains Ets motif like site (A), HOXñPBX recognition sequence (B), unknown sequence as DNA binding consensus (C), CRE (D), COUP-TFII (ARP-1) motif like site (E), and AGE3 like site (F). Footprint D has been characterized by means of functional studies as the genuine human renin gene CRE interacting with CREB in cooperation with the site of footprint B. To obtain further clues to the specific expression in the promoter region, these putative cis-elements were conducted to a consensus-specific binding assay to compare renin-producing and non-renin-producing cells by EMSA and electromobility super-shift assay. Different sequence-specific DNA/protein binding was obtained among the different cell lines with footprint B site, with COUP-TFII (ARP-1) motif like site and possibly with footprint F site. The results implicate these putative cis-elements and each corresponding trans-factor in the specific expression of the human renin gene in the promoter region. Further functional characterization of these elements would provide important data for a better understanding of human renin gene expression.
Konoshita
Tadashi
T
Third Department of Internal Medicine, Fukui University School of Medicine, Shimoaizuki, Matsuoka, Japan. kinosita@fmsrsa.fukui-med.ac.jp
Makino
Yasukazu
Y
Wakahara
Shigeyuki
S
Ido
Kanako
K
Yoshida
Masahiro
M
Kawai
Yasuyuki
Y
Miyamori
Isamu
I
D000818:Animals; D001483:Base Sequence; D051837:COUP Transcription Factor II; D051836:COUP Transcription Factors; D002457:Cell Extracts / Q000737:chemistry; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002522:Chlorocebus aethiops; D017362:Cyclic AMP Response Element-Binding Protein / Q000302:isolation & purification / Q000378:metabolism; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D009841:Oligonucleotides / Q000235:genetics; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D011987:Receptors, Steroid / Q000378:metabolism; D012083:Renin / Q000235:genetics*; D020218:Response Elements / Q000235:genetics; D016415:Sequence Alignment; D013347:Subcellular Fractions / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D051837:COUP Transcription Factor II; D051836:COUP Transcription Factors; D002457:Cell Extracts; D017362:Cyclic AMP Response Element-Binding Protein; D004268:DNA-Binding Proteins; D009841:Oligonucleotides; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D011987:Receptors, Steroid; D012083:Renin; D014157:Transcription Factors; C095569:NR2F2 protein, human