title: Transcriptional regulation of fibrillin-2 gene by E2F family members in chondrocyte differentiation.
abstract: Mutation in fibrillin-2, a major structural component of extracellular microfibrils in connective tissue, results in the autosomal dominant disease congenital contractural arachnodactyly. This genetic disease is characterized by dolichostenomelia and arachnodactyly, in addition to contractures of the large joints and abnormal pinnae formation, thus indicating the significance of fibrillin-2 in chondrogenesis. In this study, we investigated the transcriptional regulation of fibrillin-2 in chondrogenic differentiation. Although mRNA expression of fibrillin-1, a highly homologous protein to fibrillin-2, remained almost unchanged during chondrogenesis of mouse ATDC5 cells, fibrillin-2 mRNA expression varied. Fibrillin-2 was highly expressed at the early stage and declined progressively during differentiation. The 5'-flanking region of the fibrillin-2 gene contains potential binding sites for E2F, Runx, AP-2, and Sox transcription factors. The promoter activity of fibrillin-2 decreased markedly following deletion and mutagenesis of the E2F binding site between -143 and -136 bp. Overexpression of E2F1 resulted in a marked increase in its promoter activity, whereas expression of other transcription factors including AP-2alpha and Runx2 had no effect. The increase in promoter activity by E2F1 was completely suppressed by the coexpression of E2F4. E2F2 and E2F3 had positive effects on the promoter activity. Although ATDC5 cells expressed transcripts for the E2F family genes at all stages of differentiation, the expression profiles differed. E2F1 expression remained almost unchanged, whereas E2F4 expression increased markedly at the late stage of differentiation. These results indicated that coordinated expression of the E2F family is critical for the transcriptional regulation of fibrillin-2 during chondrogenesis.
authors.affiliation: Department of Biological Science and Technology, the University of Tokushima Graduate School, Tokushima, Japan.
authors.forename: Takuya
authors.initials: T
authors.lastname: Yanagino
authors.forename: Keizo
authors.initials: K
authors.lastname: Yuasa
authors.forename: Masami
authors.initials: M
authors.lastname: Nagahama
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Matsuda
authors.forename: Akihiko
authors.initials: A
authors.lastname: Tsuji
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D019902:Chondrocytes / Q000166:cytology*; D020219:Chondrogenesis / Q000235:genetics; D050684:E2F Transcription Factors / Q000502:physiology*; D050687:E2F1 Transcription Factor; D050703:E2F2 Transcription Factor; D050699:E2F3 Transcription Factor; D050704:E2F4 Transcription Factor; D000071838:Fibrillin-1; D000071840:Fibrillin-2; D000071837:Fibrillins; D005786:Gene Expression Regulation / Q000502:physiology*; D051379:Mice; D008840:Microfilament Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D014158:Transcription, Genetic; D054392:Adipokines
chemical_list: D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D050703:E2F2 Transcription Factor; D050699:E2F3 Transcription Factor; D050704:E2F4 Transcription Factor; D000071838:Fibrillin-1; D000071840:Fibrillin-2; D000071837:Fibrillins; D008840:Microfilament Proteins; D012333:RNA, Messenger; C000605724:Fbn1 protein, mouse; D054392:Adipokines; C000605728:Fbn2 protein, mouse