Regulation of osteoblast-specific factor-1 (OSF-1) mRNA expression by dual promoters as revealed by RT-PCR.
OSF-1 (osteoblast-specific factor-1), which is also referred to as p18, HBBM, HB-GAM, HBGF-8, HARP, HBNF, and pleiotrophin, is a 121-amino acid polypeptide that can induce neurite outgrowth in vitro and is highly expressed in several tissues during fetal development but exhibits expression restricted to brain and bone tissues in adults. We have reported the genomic structure of mouse OSF-1 gene, in which the open reading frame spans four exons and at least two additional 5'-UTR exons (upstream exon U2 and downstream exon U1) exist. From analysis of isolated cDNAs, two types of cDNAs were identified: one has a sequence for U1 and U2 and the other has a sequence for an intron (present between U1 and U2) and U1. This suggests that the OSF-1 gene utilizes two alternative promoters, a distal and a proximal promoter, designated promoters II and I, respectively, for the translation initiation site (ATG). Promoter II is thought to exist upstream of the intron, while promoter I is present in the intron. RT-PCR was employed to examine which OSF-1 promoters are used during development and in various cell lines. In adult mice (aged 2 months), usage of promoter I was predominant, and OSF-1 mRNAs were expressed in many organs including brain and bone. At one fetal stage (E-19), promoter I was active in the major organs including brain, liver, kidney, and intestine, while promoter II was active only in the brain. In the cell lines examined, usage of promoter I was frequent, while promoter II was active only in a few cell lines such as MC3T3-E1 (cultured for 7 days) and C3H10T1/2. These findings suggest that OSF-1 may play fundamental roles in differentiation, growth and maintenance of adult organs as well as in embryogenesis, and indicate that the expression of OSF-1 is regulated, at least in part, by the usage of different promoters in the mouse.
Sato
M
M
Molecular Medicine Research Center, The Institute of Medical Sciences, Tokai University, Kanagawa, Japan. masasato@is.icc.u-tokai.ac.jp
Miyado
K
K
Okazaki
M
M
Kimura
M
M
D000375:Aging; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D017931:DNA Primers; D004622:Embryo, Mammalian; D005260:Female; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics; D010006:Osteoblasts / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D012886:Skull / Q000378:metabolism
D002352:Carrier Proteins; D016207:Cytokines; D017931:DNA Primers; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C066845:pleiotrophin