Simultaneous Transport of Different Localized mRNA Species Revealed by Live-Cell Imaging
| Autor: | Ondrej Burkacky, Ralf-Peter Jansen, Don C. Lamb, Christoph Bräuchle, Maria Schmid, Susanne Lange, Yoshihiko Katayama |
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| Rok vydání: | 2008 |
| Předmět: |
RNA localization
Saccharomyces cerevisiae Intracellular mRNA localization Biology Models Biological Biochemistry Structural Biology Live cell imaging P-bodies Image Processing Computer-Assisted Genetics Animals Humans Bacteriophages RNA Messenger Molecular Biology Ribonucleoprotein Microscopy Confocal Lasers RNA-Binding Proteins RNA Biological Transport Cell Biology biology.organism_classification Fusion protein Cell biology Microscopy Fluorescence Ribonucleoproteins Drosophila |
| Zdroj: | Traffic. 9:1256-1267 |
| ISSN: | 1600-0854 1398-9219 |
| DOI: | 10.1111/j.1600-0854.2008.00763.x |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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