The Drosophila hairy RNA localization signal modulates the kinetics of cytoplasmic mRNA transport.
In several Drosophila cell types, mRNA transport depends on microtubules, the molecular motor dynein and trans-acting factors including Egalitarian and Bicaudal-D. However, the molecular basis of transcript recognition by the localization machinery is poorly understood. Here, we characterize the features of hairy pair-rule RNA transcripts that mediate their apical localization, using in vivo injection of fluorescently labelled mRNAs into syncytial blastoderm embryos. We show that a 121-nucleotide element within the 3'-untranslated region is necessary and sufficient to mediate apical transport. The signal comprises two essential stem-loop structures, in which double-stranded stems are crucial for localization. Base-pair identities within the stems are not essential, but can contribute to the efficiency of localization, suggesting that specificity is mediated by higher-order structure. Using time-lapse microscopy, we measure the kinetics of localization and show that impaired localization of mutant signals is due to delayed formation of active motor complexes and, unexpectedly, to slower movement. These findings, and those from co-injecting wild-type and mutant RNAs, suggest that the efficiency of molecular motors is modulated by the character of their cargoes.
Bullock
Simon L
SL
Developmental Genetics and Light Microscopy Laboratories, Cancer Research UK, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Zicha
Daniel
D
Ish-Horowicz
David
D
D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins; D016764:Cell Polarity; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics / Q000502:physiology*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D021381:Protein Transport / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D016415:Sequence Alignment
D051792:Basic Helix-Loop-Helix Proteins; D029721:Drosophila Proteins; D019476:Insect Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C061998:BicD protein, Drosophila; C104613:egl protein, Drosophila; C489235:hairy protein, Drosophila; C061841:hairy protein, insect