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Localization of a bacterial group II intron-encoded protein in human cells

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  • Mercedes Reinoso-Colacio
  • Fernando Manuel García-Rodríguez
  • Marta García-Cañadas
  • Suyapa Amador-Cubero
  • José Luis García Pérez
  • Nicolás Toro

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Original languageEnglish
Pages (from-to)12716
JournalScientific Reports
Volume5
Early online date5 Aug 2015
DOIs
Publication statusE-pub ahead of print - 5 Aug 2015

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.

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