Bazooka recruits Inscuteable to orient asymmetric cell divisions in Drosophila neuroblasts

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Abstract

Asymmetric cell divisions can be generated by the segregation of determinants into one of the two daughter cells1,2,3,4. In Drosophila, neuroblasts divide asymmetrically along the apical–basal axis shortly after their delamination from the neuroectodermal epithelium. Several proteins, including Numb and Miranda, segregate into the basal daughter cell and are needed for the determination of its correct cell fate5,6,7,8,9,10. Both the apical–basal orientation of the mitotic spindle and the localization of Numb and Miranda to the basal cell cortex are directed by Inscuteable, a protein that localizes to the apical cell cortex before and during neuroblast mitosis11,12. Here we show that the apical localizaton of Inscuteable requires Bazooka, a protein containing a PDZ domain that is essential for apical–basal polarity in epithelial cells13,14. Bazooka localizes with Inscuteable in neuroblasts and binds to the Inscuteable localization domain in vitro and in vivo. In embryos lacking both maternal and zygotic bazooka function, Inscuteable no longer localizes asymmetrically in neuroblasts and is instead uniformly distributed in the cytoplasm. Mitotic spindles in neuroblasts are misoriented in these embryos, and the proteins Numb and Miranda fail to localize asymmetrically in metaphase. Our results suggest that direct binding to Bazooka mediates the asymmetric localization of Inscuteable and connects the asymmetric division of neuroblasts to the axis of epithelial apical–basal polarity.

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Figure 1: Localization and binding of Inscuteable and Bazooka.
Figure 2: Defects in Inscuteable localization in bazooka embryos.
Figure 3: Defects in Miranda and Numb localization in the absence of Bazooka and Inscuteable.
Figure 4: Model for Bazooka and Inscuteable function during asymmetric cell division in neuroblasts.

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Acknowledgements

We thank M. Glotzer for stimulating discussions; G. Christofori, B. Dickson and M. Glotzer for helpful comments on the manuscript; E. Kleiner for technical assistance; P. Steinlein and K. Paiha for help with confocal microscopy; W. Chia and Y. N. Jan for providing antibodies; K. S. McKim and A. Müller for providing fly stocks; and A. Wodarz and E. Knust for communicating results before publication.

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Correspondence to Juergen A. Knoblich.

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Schober, M., Schaefer, M. & Knoblich, J. Bazooka recruits Inscuteable to orient asymmetric cell divisions in Drosophila neuroblasts. Nature 402, 548–551 (1999) doi:10.1038/990135

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