title: Strategies for investigating nuclear-cytoplasmic tRNA dynamics in yeast and mammalian cells.
abstract: Nuclear-cytoplasmic tRNA transport involves multiple pathways that are segregated by the involvement of distinct proteins. The tRNA export process begins in the nucleolus, where the functionality of newly produced tRNAs are tested by aminoacylation, and ends with the delivery of the exported aminoacyl tRNAs to the eukaryotic elongation factor eEF-1A for utilization in protein synthesis in the cytoplasm. Recent studies have identified a number of proteins that participate in nuclear tRNA export in both yeast and mammals. However, genetic and biochemical evidence suggest that additional components, which have yet to be identified, also participate in nuclear-cytoplasmic tRNA trafficking. Here we review key strategies that have led to the identification and characterization of proteins that are involved in the nuclear tRNA export process in yeasts and mammals. The approaches described will greatly facilitate the identification and delineation of the roles of new proteins involved in nuclear export of tRNAs to the cytoplasm.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
authors.forename: Jacqueline B
authors.initials: JB
authors.lastname: Pierce
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
authors.forename: Shawn C
authors.initials: SC
authors.lastname: Chafe
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
authors.forename: Manoja B K
authors.initials: MB
authors.lastname: Eswara
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
authors.forename: George
authors.initials: G
authors.lastname: van der Merwe
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada.
authors.forename: Dev
authors.initials: D
authors.lastname: Mangroo
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000502:physiology*; D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D017930:Genes, Reporter / Q000235:genetics; D016153:Genes, Suppressor; D006367:HeLa Cells; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D028884:Karyopherins / Q000235:genetics; D028861:Nuclear Pore Complex Proteins / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D009697:Nucleolus Organizer Region / Q000235:genetics*; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D034443:RNA Transport / Q000502:physiology*; D012343:RNA, Transfer / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D046249:Transfer RNA Aminoacylation / Q000502:physiology*; D020798:Two-Hybrid System Techniques; D028961:beta Karyopherins / Q000378:metabolism
chemical_list: C094814:EEF1A1 protein, human; D028884:Karyopherins; C408090:Los1 protein, S cerevisiae; C401306:MSN5 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D020648:Peptide Elongation Factor 1; D016601:RNA-Binding Proteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; C588166:Utp22 protein, S cerevisiae; C524393:Utp8 protein, S cerevisiae; C547588:Utp9 protein, S cerevisiae; D028961:beta Karyopherins; D012343:RNA, Transfer
keywords: Fluorescence in situ hybridization; In vivo nuclear tRNA export assay; Mammalian cells; Nuclear tRNA export factors; Separation of aminoacylated and nonaminoacylated tRNA; Yeast; tRNA three-hybrid interaction