Selection of homeotic proteins for binding to a human DNA replication origin
| Autor: | E. d. Stanchina, D. Gabellini, P. Norio, F. A. Peverali, S. Riva, A. Falaschi, G. Biamonti, GIACCA, MAURO |
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| Přispěvatelé: | E. d., Stanchina, D., Gabellini, P., Norio, Giacca, Mauro, F. A., Peverali, S., Riva, A., Falaschi, G., Biamonti |
| Jazyk: | angličtina |
| Rok vydání: | 2000 |
| Předmět: |
chemistry/genetics/metabolism
Humans Lamin Type B Lamins Mice Molecular Sequence Data Nuclear Protein Recombinant Fusion Proteins Molecular Sequence Data Messenger DNA Footprinting genetics/metabolism genetics Homeodomain Protein Replication Origin analysis/genetics analysis/genetics Recombinant Fusion Protein Transfection genetics/metabolism Gene Expression Regulation Gene Cell Line Substrate Specificity Mice Open Reading Frames genetics Substrate Specificity Transfection Two-Hybrid System Techniques Two-Hybrid System Techniques genetics Organ Specificity Protein Binding RNA Animals Homeobox Humans genetics RNA Messenger Amino Acid Sequence Animals Base Sequence Cell Division Cell Line DNA Footprinting DNA genetics/metabolism Gene Expression Regulation Genes genetics Homeodomain Proteins chemistry/genetics/metabolism Humans Lamin Type B Lamins Mice Molecular Sequence Data Nuclear Proteins genetics Open Reading Frames analysis/genetics Recombinant Fusion Proteins chemistry/genetics/metabolism Replication Origin Amino Acid Sequence chemistry/genetics/metabolism Homeodomain Proteins Base Sequence Lamin Type B Genes Homeobox Nuclear Proteins DNA genetics Open Reading Frame Lamins Gene Expression Regulation Genes Organ Specificity RNA Cell Division Protein Binding |
| Popis: | We have previously shown that a cell cycle-dependent nucleoprotein complex assembles in vivo on a 74 bp sequence within the human DNA replication origin associated to the Lamin B2 gene. Here, we report the identification, using a one-hybrid screen in yeast, of three proteins interacting with the 74 bp sequence. All of them, namely HOXA13, HOXC10 and HOXC13, are orthologues of the Abdominal-B gene of Drosophila melanogaster and are members of the homeogene family of developmental regulators. We describe the complete open reading frame sequence of HOXC10 and HOXC13 along with the structure of the HoxC13 gene. The specificity of binding of these two proteins to the Lamin B2 origin is confirmed by both band-shift and in vitro footprinting assays. In addition, the ability of HOXC10 and HOXC13 to increase the activity of a promoter containing the 74 bp sequence, as assayed by CAT-assay experiments, demonstrates a direct interaction of these homeoproteins with the origin sequence in mammalian cells. We also show that HOXC10 expression is cell-type-dependent and positively correlates with cell proliferation. |
| Databáze: | OpenAIRE |
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