title: In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p.
abstract: Many messenger RNA export proteins have been identified; yet the spatial and temporal activities of these proteins and how they determine directionality of messenger ribonucleoprotein (mRNP) complex export from the nucleus remain largely undefined. Here, the bacteriophage PP7 RNA-labeling system was used in Saccharomyces cerevisiae to follow single-particle mRNP export events with high spatial precision and temporal resolution. These data reveal that mRNP export, consisting of nuclear docking, transport, and cytoplasmic release from a nuclear pore complex (NPC), is fast (∼ 200 ms) and that upon arrival in the cytoplasm, mRNPs are frequently confined near the nuclear envelope. Mex67p functions as the principal mRNP export receptor in budding yeast. In a mex67-5 mutant, delayed cytoplasmic release from NPCs and retrograde transport of mRNPs was observed. This proves an essential role for Mex67p in cytoplasmic mRNP release and directionality of transport.
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: Carlas
authors.initials: C
authors.lastname: Smith
authors.affiliation: Department of Cell Biology, University of Alberta, T6G 2H7 Edmonton, Alberta, Canada.
authors.forename: Azra
authors.initials: A
authors.lastname: Lari
authors.affiliation: Department of Biology, Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Carina Patrizia
authors.initials: CP
authors.lastname: Derrer
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605 Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
authors.forename: Anette
authors.initials: A
authors.lastname: Ouwehand
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605 Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
authors.forename: Ammeret
authors.initials: A
authors.lastname: Rossouw
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605 Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
authors.forename: Maximiliaan
authors.initials: M
authors.lastname: Huisman
authors.affiliation: Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
authors.forename: Thomas
authors.initials: T
authors.lastname: Dange
authors.affiliation: Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
authors.forename: Mark
authors.initials: M
authors.lastname: Hopman
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605.
authors.forename: Aviva
authors.initials: A
authors.lastname: Joseph
authors.affiliation: Departement de Biochimie et Medecine Moleculaire, Universite de Montreal, H3T 1J4 Montreal, Quebec, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Zenklusen
authors.affiliation: Department of Biology, Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland Department of Cell and Developmental Biology, University of California, Berkeley, Berkeley, CA 94720 ben.montpetit@ualberta.ca david.grunwald@umassmed.edu karsten.weis@bc.biol.ethz.ch.
authors.forename: Karsten
authors.initials: K
authors.lastname: Weis
authors.affiliation: RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605 Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands ben.montpetit@ualberta.ca david.grunwald@umassmed.edu karsten.weis@bc.biol.ethz.ch.
authors.forename: David
authors.initials: D
authors.lastname: Grunwald
authors.affiliation: Department of Cell Biology, University of Alberta, T6G 2H7 Edmonton, Alberta, Canada ben.montpetit@ualberta.ca david.grunwald@umassmed.edu karsten.weis@bc.biol.ethz.ch.
authors.forename: Ben
authors.initials: B
authors.lastname: Montpetit
mesh_terms: D002467:Cell Nucleus / Q000235:genetics* / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism*; D034443:RNA Transport; D012331:RNA, Fungal / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: C106898:MEX67 protein, S cerevisiae; D009687:Nuclear Proteins; D029543:Nucleocytoplasmic Transport Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029701:Saccharomyces cerevisiae Proteins