title: Localization of a bacterial group II intron-encoded protein in human cells.
abstract: Group II introns are mobile retroelements that self-splice from precursor RNAs to form ribonucleoparticles (RNP), which can invade new specific genomic DNA sites. This specificity can be reprogrammed, for insertion into any desired DNA site, making these introns useful tools for bacterial genetic engineering. However, previous studies have suggested that these elements may function inefficiently in eukaryotes. We investigated the subcellular distribution, in cultured human cells, of the protein encoded by the group II intron RmInt1 (IEP) and several mutants. We created fusions with yellow fluorescent protein (YFP) and with a FLAG epitope. We found that the IEP was localized in the nucleus and nucleolus of the cells. Remarkably, it also accumulated at the periphery of the nuclear matrix. We were also able to identify spliced lariat intron RNA, which co-immunoprecipitated with the IEP, suggesting that functional RmInt1 RNPs can be assembled in cultured human cells.
authors.affiliation: Grupo de Ecología Genética, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Calle Profesor Albareda 1, 18008 Granada, Spain.
authors.forename: Mercedes
authors.initials: M
authors.lastname: Reinoso-Colacio
authors.affiliation: Grupo de Ecología Genética, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Calle Profesor Albareda 1, 18008 Granada, Spain.
authors.forename: Fernando Manuel
authors.initials: FM
authors.lastname: García-Rodríguez
authors.affiliation: GENYO (Centro Pfizer-Universidad de Granada-Junta de Andalucía de Genómica e Investigación Oncológica), PTS Granada, Avda. de la Ilustración 114 18016 Granada, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: García-Cañadas
authors.affiliation: GENYO (Centro Pfizer-Universidad de Granada-Junta de Andalucía de Genómica e Investigación Oncológica), PTS Granada, Avda. de la Ilustración 114 18016 Granada, Spain.
authors.forename: Suyapa
authors.initials: S
authors.lastname: Amador-Cubero
authors.affiliation: GENYO (Centro Pfizer-Universidad de Granada-Junta de Andalucía de Genómica e Investigación Oncológica), PTS Granada, Avda. de la Ilustración 114 18016 Granada, Spain.
authors.forename: José Luis
authors.initials: JL
authors.lastname: García Pérez
authors.affiliation: Grupo de Ecología Genética, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Calle Profesor Albareda 1, 18008 Granada, Spain.
authors.forename: Nicolás
authors.initials: N
authors.lastname: Toro
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology; D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D019556:COS Cells; D002522:Chlorocebus aethiops; D006367:HeLa Cells; D006801:Humans; D007438:Introns / Q000502:physiology*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D012326:RNA Splicing / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012261:Ribonucleoproteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; C054562:yellow fluorescent protein, Bacteria