Molecular cloning, developmental expression, promoter analysis and functional characterization of the mouse CNBP gene.
Striking conservation in various organisms suggests that cellular nucleic acid-binding protein (CNBP) plays a fundamental biological role across different species. However, the regulated expression and physiological properties of the CNBP gene are unknown. In this study, we report the molecular cloning, promoter characterization, developmental expression and functional analysis of the mouse CNBP gene. The gene contains five exons and is localized to chromosome 6 in the region corresponding to band 6 D1-D2. Primer extension assay indicates that the transcription start site is located 230 bp upstream of the initiator Met codon. Our promoter analysis indicates that strong transcription enhancer and silencer regions lie within the 1.6 kb proximal region of the promoter and the upstream -3.0 to -1.6 kb region, respectively. The promoter activity is 10 fold higher in embryonic carcinoma cells than that in fibroblast, as determined by CAT assay. Consistent with its function as a transcription factor, CNBP protein is located in the nucleus of cells. During mouse embryogenesis, CNBP is expressed in the anterior region of the early embryo and in the limb, tail and craniofacial region. Overexpression of CNBP strongly stimulates cell proliferation and increases c-myc promoter activity. Our finds suggest that CNBP may play an important role in cell proliferation and tissue patterning during anterior-posterior axis, craniofacial and limb development by targeting c-Myc.
Shimizu
Ken
K
Department of Cytokine Biology, The Forsyth Institute, Boston, MA 02115, USA.
Chen
Wei
W
Ashique
Amir M
AM
Moroi
Ryoji
R
Li
Yi-Ping
YP
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D005091:Exons; D018507:Gene Expression Regulation, Developmental; D005796:Genes / Q000235:genetics; D017403:In Situ Hybridization; D017404:In Situ Hybridization, Fluorescence; D007438:Introns; D051379:Mice; D008809:Mice, Inbred C3H; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D016601:RNA-Binding Proteins; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D024363:Transcription Initiation Site; D014162:Transfection; D014407:Tumor Cells, Cultured
D015500:Chloramphenicol O-Acetyltransferase; D004268:DNA-Binding Proteins; D016271:Proto-Oncogene Proteins c-myc; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C499018:Cnbp protein, rat; C499012:Cnbp protein, mouse