Miranda mediates asymmetric protein and RNA localization in the developing nervous system.
Neuroblasts undergo asymmetric stem cell divisions to generate a series of ganglion mother cells (GMCs). During these divisions, the cell fate determinant Prospero is asymmetrically partitioned to the GMC by Miranda protein, which tethers it to the basal cortex of the dividing neuroblast. Interestingly, prospero mRNA is similarly segregated by the dsRNA binding protein, Staufen. Here we show that Staufen interacts in vivo with a segment of the prospero 3' UTR. Staufen protein and prospero RNA colocalize to the apical side of the neuroblast at interphase, but move to the basal side during prophase. Both the apical and basal localization of Staufen are abolished by the removal of a conserved domain from the carboxyl terminus of the protein, which interacts in a yeast two-hybrid screen with Miranda protein. Furthermore, Miranda colocalizes with Staufen protein and prospero mRNA during neuroblast divisions, and neither Staufen nor prospero RNA are localized in miranda mutants. Thus Miranda, which localizes Prospero protein, also localizes prospero RNA through its interaction with Staufen protein.
Schuldt
A J
AJ
Wellcome/CRC Institute and Department of Genetics, Cambridge CB2 1QR, UK.
Adams
J H
JH
Davidson
C M
CM
Micklem
D R
DR
Haseloff
J
J
St Johnston
D
D
Brand
A H
AH
D000818:Animals; D001665:Binding Sites / Q000502:physiology; D002453:Cell Cycle / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002467:Cell Nucleus / Q000737:chemistry / Q000502:physiology; D003593:Cytoplasm / Q000737:chemistry / Q000502:physiology; D004330:Drosophila / Q000737:chemistry / Q000196:embryology / Q000502:physiology; D029721:Drosophila Proteins; D017952:Ganglia, Invertebrate / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D019476:Insect Proteins / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000235:genetics; D009420:Nervous System / Q000737:chemistry / Q000166:cytology* / Q000196:embryology*; D009687:Nuclear Proteins / Q000235:genetics; D011485:Protein Binding; D012313:RNA / Q000032:analysis / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D015534:Trans-Activators / Q000502:physiology; D014157:Transcription Factors
D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; D019476:Insect Proteins; C107613:Mira protein, Drosophila; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C073099:pros protein, Drosophila; C069927:stau protein, Drosophila; D012313:RNA