Simultaneous Transport of Different Localized mRNA Species Revealed by Live-Cell Imaging

Authors

  • Susanne Lange,

    1. Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, D-81377 Munich, Germany
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  • Yoshihiko Katayama,

    1. Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstrasse 11, D-81377 Munich, Germany
    2. Center for Nanoscience (CeNS), Geschwister-Scholl-Platzl, 80539 Munich, Germany
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  • Maria Schmid,

    1. Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, D-81377 Munich, Germany
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  • Ondrej Burkacky,

    1. Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstrasse 11, D-81377 Munich, Germany
    2. Center for Nanoscience (CeNS), Geschwister-Scholl-Platzl, 80539 Munich, Germany
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  • Christoph Bräuchle,

    1. Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstrasse 11, D-81377 Munich, Germany
    2. Center for Nanoscience (CeNS), Geschwister-Scholl-Platzl, 80539 Munich, Germany
    3. Munich Center for Integrated Protein Science (CiPSM), Ludwig-Maximilians-Universität München, Butenandtstrasse 5-13, D-81377 Munich, Germany
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  • Don C. Lamb,

    Corresponding author
    1. Department of Chemistry and Biochemistry, Ludwig-Maximilians-Universität München, Butenandtstrasse 11, D-81377 Munich, Germany
    2. Center for Nanoscience (CeNS), Geschwister-Scholl-Platzl, 80539 Munich, Germany
    3. Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
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  • Ralf-Peter Jansen

    Corresponding author
    1. Gene Center, Ludwig-Maximilians-Universität München, Feodor-Lynen-Str. 25, D-81377 Munich, Germany
    2. Munich Center for Integrated Protein Science (CiPSM), Ludwig-Maximilians-Universität München, Butenandtstrasse 5-13, D-81377 Munich, Germany
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*Ralf-Peter Jansen, rjansen@lmb.uni-muenchen.de and Don C. Lamb, don.lamb@cup.uni-muenchen.de

Abstract

Intracellular mRNA localization is a common mechanism to achieve asymmetric distributions of proteins. Previous studies have revealed that in a number of cell types, different mRNA species are localized by the same transport machinery. However, it has been unclear if these individual mRNA species are specifically sorted into separate or common ribonucleoprotein (RNP) particles before or during transport. Using budding yeast as a model system, we analyzed the intracellular movement of individual pairs of localized mRNA in live cells. Yeast cells localize more than 20 different mRNAs to the bud with the help of the Myo4p/She3p/She2p protein complex. For live cell imaging, mRNA pairs were tagged with tandem repeats of either bacteriophage MS2 or lambda boxB RNA sequences and fluorescently labeled by fusion protein constructs that bind to the RNA tag sequences. Using three-dimensional, single-particle tracking with dual-color detection, we have tracked the transport of two different localized mRNA species in real time. Our observations show that different localized mRNAs are coassembled into common RNP particles and cotransported in a directional manner to the target site. Nonlocalized mRNAs or mutant mRNAs that lack functional localization signals form separate particles that are not transported to the bud. This study reveals a high degree of co-ordination of mRNA trafficking in budding yeast.

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