A functional analysis of inscuteable and its roles during Drosophila asymmetric cell divisions.
Cellular diversity in the Drosophila central nervous system is generated through a series of asymmetric cell divisions in which one progenitor produces two daughter cells with distinct fates. Asymmetric basal cortical localisation and segregation of the determinant Prospero during neuroblast cell divisions play a crucial role in effecting distinct cell fates for the progeny sibling neuroblast and ganglion mother cell. Similarly asymmetric localisation and segregation of the determinant Numb during ganglion mother cell divisions ensure that the progeny sibling neurons attain distinct fates. The most upstream component identified so far which acts to organise both neuroblast and ganglion mother cell asymmetric divisions is encoded by inscuteable. The Inscuteable protein is itself asymmetrically localised to the apical cell cortex and is required both for the basal localisation of the cell fate determinants during mitosis and for the orientation of the mitotic spindle along the apical/basal axis. Here we define the functional domains of Inscuteable. We show that aa252-578 appear sufficient to effect all aspects of its function, however, the precise requirements for its various functions differ. The region, aa288-497, is necessary and sufficient for apical cortical localisation and for mitotic spindle (re)orientation along the apical/basal axis. A larger region aa288-540 is necessary and sufficient for asymmetric Numb localisation and segregation; however, correct localisation of Miranda and Prospero requires additional sequences from aa540-578. The requirement for the resolution of distinct sibling neuronal fates appears to coincide with the region necessary and sufficient for Numb localisation (aa288-540). Our data suggest that apical localisation of the Inscuteable protein is a necessary prerequisite for all other aspects of its function. Finally, we show that although inscuteable RNA is normally apically localised, RNA localisation is not required for protein localisation or any aspects of inscuteable function.
Tio
M
M
Institute of Molecular and Cell Biology, National University of Singapore Campus, Singapore 117609.
Zavortink
M
M
Yang
X
X
Chia
W
W
D000818:Animals; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000378:metabolism; D002455:Cell Division; D003598:Cytoskeletal Proteins / Q000378:metabolism / Q000502:physiology*; D004330:Drosophila / Q000196:embryology*; D029721:Drosophila Proteins; D005455:Fluorescent Antibody Technique; D007605:Juvenile Hormones / Q000378:metabolism; D008957:Models, Genetic; D016296:Mutagenesis; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009479:Neuropeptides; D009687:Nuclear Proteins / Q000378:metabolism; D013696:Temperature; D014157:Transcription Factors
D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D029721:Drosophila Proteins; D007605:Juvenile Hormones; C107613:Mira protein, Drosophila; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D009687:Nuclear Proteins; D014157:Transcription Factors; C098678:insc protein, Drosophila; C060606:numb protein, Drosophila; C073099:pros protein, Drosophila