In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p

Carlas Smith; Azra Lari; Carina Patrizia Derrer; Anette Ouwehand; Ammeret Rossouw; Maximiliaan Huisman; Thomas Dange; Mark Hopman; Aviva Joseph; Daniel Zenklusen; Karsten Weis; David Grunwald; Ben Montpetit

doi:10.1083/jcb.201503135

In vivo single-particle imaging of nuclear mRNA export in budding yeast demonstrates an essential role for Mex67p

J Cell Biol. 2015 Dec 21;211(6):1121-30. doi: 10.1083/jcb.201503135.

Authors

Affiliations

¹ RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605 Department Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA 01605.
² Department of Cell Biology, University of Alberta, T6G 2H7 Edmonton, Alberta, Canada.
³ Department of Biology, Institute of Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
⁴ 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.
⁵ Bionanoscience, Kavli Institute of Nanoscience, Delft University of Technology, 2628 CJ Delft, Netherlands.
⁶ Departement de Biochimie et Medecine Moleculaire, Universite de Montreal, H3T 1J4 Montreal, Quebec, Canada.
⁷ 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.
⁸ 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.
⁹ 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.

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.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Nucleus / genetics*
Cell Nucleus / metabolism*
Nuclear Proteins / metabolism*
Nucleocytoplasmic Transport Proteins / metabolism*
RNA Transport*
RNA, Fungal / metabolism
RNA, Messenger / metabolism*
RNA-Binding Proteins / metabolism*
Saccharomyces cerevisiae / genetics*
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / metabolism*

Substances

MEX67 protein, S cerevisiae
Nuclear Proteins
Nucleocytoplasmic Transport Proteins
RNA, Fungal
RNA, Messenger
RNA-Binding Proteins
Saccharomyces cerevisiae Proteins

Abstract

Publication types

MeSH terms

Substances

Grant support