Analysis of the promoter region of human placenta-specific DSCR4 gene.
The gene DSCR4 locates in the band q22.2 of human chromosome 21 and encodes a protein of 118 amino acids. Expression of DSCR4 is restricted to human placenta and placental choriocarcinoma cell lines BeWo and JEG3. The 5'-RACE method using RNA from human placenta indicated the major transcription start site at 93 nt upstream (nt -93) of the initiation codon. Transfection assay using a series of deletion constructs of the 5'-flanking region fused to the luciferase reporter gene identified three positive regions nt -2200 to -2088, nt -2064 to -1924, nt -810 to -632 and two negative regions nt -1923 to -1740, nt -631 to -425. The computer analysis predicted the presence of several cis-elements in these regions and the promoter assay using various mutants of consensus sequence identified two distinct cis-elements for OLF-1 and E47. The electrophoretic mobility shift assay (EMSA) using the extracts of DSCR4-expressing cells confirmed the binding of certain protein factors to these cis-elements. In fact, OLF-1-like transcription factor, EBF-3 and EBF-4 were detected in the DSCR4-expressing cell lines and human placenta. Based on these data, we postulated that transcription of DSCR4 gene is regulated positively by binding of OLF-1-like transcription factor and negatively by binding of E47-like transcription factor.
Asai
Satoko
S
Department of Medical Genetics, School of Health Sciences, Kyorin University, 476, Miyashita-cho, Hachioji-shi, Tokyo 192-8508, Japan.
Yamaki
Akiko
A
Kudoh
Jun
J
Shimizu
Nobuyoshi
N
Shimizu
Yoshiko
Y
D024506:5' Flanking Region; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010920:Placenta / Q000378:metabolism*; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D062085:RNA, Long Noncoding; D051820:TCF Transcription Factors / Q000378:metabolism; D014018:Tissue Distribution; D015534:Trans-Activators / Q000378:metabolism; D058125:Transcription Factor 7-Like 1 Protein; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; C110524:DSCR4 lncRNA, human; C081976:EBF1 protein, human; D011257:Pregnancy Proteins; D062085:RNA, Long Noncoding; D051820:TCF Transcription Factors; C544912:TCF7L1 protein, human; D015534:Trans-Activators; D058125:Transcription Factor 7-Like 1 Protein; D004247:DNA