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Vol. 12, Issue 9, 2567-2577, September 2001

VIDEO ESSAY
ASH1 mRNA Localization in Three Acts

Dale L. Beach,^* and Kerry Bloom

Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599-3280

Novel green fluorescent protein (GFP) labeling techniques targeting specific mRNA transcripts reveal discrete phases of mRNA localization in yeast: packaging, transport, and docking. In budding yeast, ASH1 mRNA is translocated via actin and myosin to the tip of growing cells. A GFP-decorated reporter transcript containing the ASH1 3' untranslated region gRNA_ASH1 forms spots of fluorescence localized to a cortical domain at the bud tip, relocates to the mother-bud neck before cell separation, and finally migrates to the incipient bud site before the next budding cycle. The correct positioning of the mRNA requires at least six proteins: She1p-5p and Bud6p/Aip3p. gRNA_ASH1 localization in mutant strains identified three functional categories for the She proteins: mRNA particle formation (She2p and She4p), mRNA transport into the bud (She1p/Myo4p and She3p), and mRNA tethering at the bud tip (She5p/Bni1p and Bud6p/Aip3p). Because localization of the mRNA within the bud does not a priori restrict the translated protein, we examine the distribution of a mother-specific protein (Yta6p) translated from a mRNA directed into the bud. Yta6p remains associated with the mother cortex despite localization of the mRNA to the bud. This video essay traces the life history of a localized mRNA transcript, describes the roles of proteins required to polarize and anchor the mRNA, and demonstrates at least one instance where mRNA localization does not effect protein localization.

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Online version of this article contains video material for Figures 2-7. Online version available at www.molbiolcell.org.

^* Corresponding author. E-mail address: dbeach{at}emailunc.edu.

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Molecular Biology of the Cell
Vol. 12, 2567-2577, September 2001
Copyright © 2001 by The American Society for Cell Biology

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