Cloning and characterization of the 5'-flanking region of the mouse diastrophic dysplasia sulfate transporter gene.
Dyastrophic dysplasia sulfate transporter (DTDST) plays an important role in proteoglycan synthesis in the extracellular matrix of bone and cartilage. Recently, we found that the mouse DTDST gene was induced in pluripotent C3H10T1/2 cells during differentiation by bone morphogenetic protein-2 (BMP-2). To clarify the transcriptional regulation of the DTDST gene, we have cloned the 5'-flanking region of the mouse DTDST gene by the PCR based gene walking method. Sequence analysis revealed the presence of the TATA box followed by GC rich sequences containing two Sp-1 binding sites and a CBFA1 binding site. Transient transfection assays demonstrated that the basal transcriptional activity in osteoblastic MC3T3-E1 cells was mainly present between -309 and -275 bp upstream of the transcription start site (Segment -309/-275) which contained the consensus sequence for the xenobiotic-responsible element (XRE). Nuclear proteins from MC3T3-E1 cells and C3H10T1/2 cells could bind to this short segment in vitro. BMP-2 increased the promoter activity as well as the nuclear protein binding to the sequence in C3H10T1/2 cells. The present data suggest that the DTDST gene expression in osteoblasts and differentiating precursor cells to osteoblast/chondrocyte lineage would be mainly regulated by undetermined XRE binding transcription factors.
Kobayashi
T
T
Department of Medicine, Kobe University School of Medicine, Japan.
Sugimoto
T
T
Saijoh
K
K
Fujii
M
M
Chihara
K
K
D000818:Animals; D027321:Anion Transport Proteins; D001483:Base Sequence; D001692:Biological Transport / Q000235:genetics; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular; D005347:Fibroblasts; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism; D010006:Osteoblasts; D010009:Osteochondrodysplasias / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000502:physiology; D000076988:Sulfate Transporters; D013431:Sulfates / Q000378:metabolism*
D027321:Anion Transport Proteins; D002352:Carrier Proteins; D009687:Nuclear Proteins; C513264:Slc26a2 protein, mouse; D000076988:Sulfate Transporters; D013431:Sulfates