A potential role for the nucleolus in L1 retrotransposition
| Autor: | Eric M. Ostertag, Haig H. Kazazian, Maria del Carmen Seleme, Kurt A. Engleka, John L. Goodier |
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| Rok vydání: | 2004 |
| Předmět: |
Small RNA
Cytoplasm Umbilical Veins Nucleolus Immunocytochemistry Green Fluorescent Proteins Nuclear Localization Signals Cell Culture Techniques Biology Green fluorescent protein Viral Proteins Genetics medicine T7 RNA polymerase Humans Cloning Molecular Molecular Biology Gene Genetics (clinical) Immunochemistry Endothelial Cells RNA-Directed DNA Polymerase General Medicine DNA-Directed RNA Polymerases Subcellular localization Endonucleases Molecular biology Long Interspersed Nucleotide Elements Ribonucleoproteins DNA Transposable Elements Cell Nucleolus medicine.drug |
| Zdroj: | Human molecular genetics. 13(10) |
| ISSN: | 0964-6906 |
| Popis: | Determining the subcellular localization of the L1 ORF2 protein (ORF2p) has been impossible to date because of technical limitations in detecting either endogenous or overexpressed forms of the protein. Here we report visualization of the full-length ORF2p in cultured human cells following expression in a modified vaccinia virus/T7 RNA polymerase (MVA/T7RP) system. The MVA/T7RP system was used to ascertain subcellular localization of L1 ORF1p and ORF2p both as fusions with green fluorescent protein and by immunocytochemistry. Full-length ORF2p was predominantly cytoplasmic, while carboxy-terminal-deleted ORF2p localized additionally to the nucleolus. We mapped a functional nucleolar localization signal in ORF2p. ORF1p appeared in the cytoplasm with a speckled pattern and colocalized with ORF2p in nucleoli in a subset of cells. These findings help explain the presence of chimeras between L1s and small RNA gene sequences recently discovered in the human genome. |
| Databáze: | OpenAIRE |
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