Cloning and characterization of the promoter region of the mouse frizzled-related protein 4 gene.
Frizzled-related protein (Frp) is a newly identified family of secreted proteins involved in the Wnt signaling pathway. To date, little is known about the underlying mechanisms regulating Frp expression. In this study the promoter region of mouse frizzled related protein 4 (sFrp4) gene was cloned, sequenced, and analyzed using transient reporter assays along with site-directed mutagenesis. Two clusters of cis-acting elements, STAT3/Lyf-1/MZF1 (site 1) and C/EBP-beta/ GATA-1/CREB (site 2) located in the promoter region from -238 to -144 were found to be essential for the promoter activity of sFrp4. In addition to sites 1 and 2, putative transcriptional factor binding sites for TFIID, SP1/GC and ATF/CREB exhibited positive, while the site for NRSE exhibited negative regulatory functions, as determined by the alkaline phosphatase activities of the reporter assay. We also demonstrate that the ATF/CREB site may cooperatively interact with the NRSF-like element in regulating sFrp4 promoter activity. The data of our study, which is the first promoter analysis of mouse Frp genes, provide the basis for understanding the functions and the regulation of Frp and its role in regulating Wnt signals.
Wong
Vincent Kam Wai
VK
Department of Biology, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China.
Yam
Judy Wai Ping
JW
Hsiao
W L Wendy
WL
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA; D005786:Gene Expression Regulation; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics*; D051381:Rats; D016415:Sequence Alignment; D015398:Signal Transduction
D011506:Proteins; D004247:DNA