title: A simplified vector system for visualization of localized RNAs in Schizosaccharomyces pombe.
abstract: RNA localization is an important event that is essential for the polarization and differentiation of a cell. Although several methods are currently used to detect localized RNAs, a simplified detection system has not yet been developed for Schizosaccharomyces pombe. In the present study, we describe a new vector system for the visualization of localized RNAs in S. pombe using a U1A-tag-GFP system. A pREP1-U1A-tag vector plasmid to express U1A-tagged RNA and a pREP2-U1AGFP plasmid to produce a U1A-GFP fusion protein were constructed for this system. Since the U1A-GFP protein binds U1A-tagged RNA, fluorescence is observed at the location of U1A-tagged RNA in cells expressing both of these. The nucleolar localization of U3 snoRNA was successfully detected using this system, and a novel RNA localized at the DNA region of the nucleus was found by screening localized RNAs. This system will accelerate the study of localized RNAs in S. pombe.
authors.affiliation: a Junior College Division , Otsuma Women's University , Tokyo , Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takeuchi-Andoh
authors.affiliation: b Department of Biological Sciences, Graduate School of Science and Technology , Kumamoto University , Kumamoto , Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Ohba
authors.affiliation: a Junior College Division , Otsuma Women's University , Tokyo , Japan.
authors.forename: Yu
authors.initials: Y
authors.lastname: Shinoda
authors.affiliation: b Department of Biological Sciences, Graduate School of Science and Technology , Kumamoto University , Kumamoto , Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Fuchita
authors.affiliation: b Department of Biological Sciences, Graduate School of Science and Technology , Kumamoto University , Kumamoto , Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: a Junior College Division , Otsuma Women's University , Tokyo , Japan.
authors.forename: Emi
authors.initials: E
authors.lastname: Nishiyoshi
authors.affiliation: a Junior College Division , Otsuma Women's University , Tokyo , Japan.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Terouchi
authors.affiliation: b Department of Biological Sciences, Graduate School of Science and Technology , Kumamoto University , Kumamoto , Japan.
authors.forename: Tokio
authors.initials: T
authors.lastname: Tani
mesh_terms: D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D061848:Optical Imaging; D012331:RNA, Fungal / Q000235:genetics* / Q000378:metabolism; D020537:RNA, Small Nucleolar / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000235:genetics / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D012331:RNA, Fungal; D020537:RNA, Small Nucleolar; C117378:RNA, U3 small nucleolar; D011993:Recombinant Fusion Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C075339:U1A protein; D049452:Green Fluorescent Proteins
keywords: RNA localization; S. pombe; nucleus; visualization