Analysis of RNA export

Publisher Summary Nucleocytoplasmic transport plays a critical role in the expression of genetic information in eukaryotic cells. All RNAs except those encoded by the mitochondrial genome are synthesized in the nucleus, but most of these RNAs must be exported to the cytoplasm where they function in protein synthesis. The application of fluorescence in situ hybridization (FISH) to detect RNA in yeast has been critical for the analysis of RNA export. This technique is capable of providing information about the subcellular distribution of both total mRNA and individual mRNA species. Although not as quantitative as fractionation, FISH analysis also provides information about the distribution of RNA within the cytoplasmic and nuclear compartments. Because different mRNA molecules can have distinct subcellular distributions, it is sometimes useful to detect the location of specific mRNA species and—at the same time—the location of total mRNA. The ability to localize simultaneously both RNA and protein has also increased the understanding of RNA export. Finally, FISH analysis can be harnessed as a screen for mutants defective for RNA export or for distribution of a specific mRNA to its particular subcellular location.