title: The Sp1 and CBF/NF-Y transcription factors cooperatively regulate the mouse pro-alpha3(V) collagen gene (Col5a3) in osteoblastic cells.
abstract: The purpose of this study was to clarify the mechanism responsible for the transcriptional regulation of the mouse Col5a3 gene in osteoblastic cells. Transient transfection into rat osteosarcoma ROS17/2.8 cells demonstrated that a region from nucleotides 337 to 1 was involved in the transcriptional activity of the Col5a3 gene. An electrophoretic mobility shift assay showed that Sp1/Sp3 and CBF/NF-Y bound to a GC-rich domain (194/186) and a CCAAT box (134/130) in the Col5a3 gene, respectively. Introduction of mutations or deletion into a GC-rich domain, the CCAAT box, or both elements decreased the transcription activity. Overexpression of Sp1 increases the transcription activity and interferes with Sp family binding to the GC-rich domain to decrease promoter activity. Therefore, the transcription of the mouse Col5a3 gene is cooperatively regulated by Sp1 and CBF/NF-Y in osteoblastic cells.
authors.affiliation: Department of Matrix Medicine, Faculty of Medicine, Oita University, Yufu, Oita 879-5593, Japan.
authors.forename: Yun-Feng
authors.initials: YF
authors.lastname: Wu
authors.forename: Noritaka
authors.initials: N
authors.lastname: Matsuo
authors.forename: Hideaki
authors.initials: H
authors.lastname: Sumiyoshi
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Yoshioka
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D023081:CCAAT-Binding Factor / Q000502:physiology*; D002478:Cells, Cultured; D003094:Collagen / Q000235:genetics*; D005786:Gene Expression Regulation; D051379:Mice; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011498:Protein Precursors / Q000235:genetics*; D051381:Rats; D016329:Sp1 Transcription Factor / Q000502:physiology*; D024363:Transcription Initiation Site
chemical_list: D023081:CCAAT-Binding Factor; D011498:Protein Precursors; D016329:Sp1 Transcription Factor; C406808:pro-alpha3(V) collagen; D003094:Collagen