ASPM and CITK regulate spindle orientation by affecting the dynamics of astral microtubules. (25th August 2016)

• Record Type: Journal Article
• Authors: Gai, Marta; Bianchi, Federico T; Vagnoni, Cristiana; Vernì, Fiammetta; Bonaccorsi, Silvia; Pasquero, Selina; Berto, Gaia E; Sgrò, Francesco; Chiotto, Alessandra MA; Annaratone, Laura; Sapino, Anna; Bergo, Anna; Landsberger, Nicoletta; Bond, Jacqueline; Huttner, Wieland B; Di Cunto, Ferdinando
• Is Part Of: EMBO reports. Volume 17:Number 10(2016:Oct.)
• Page Start: 1396
• Page End: 1409
• Publication Date: 2016-08-25
• Subjects: astral microtubules -- asymmetric division -- microcephaly -- mitotic spindle orientation -- neurogenesis; Molecular biology -- Periodicals; Molecular Biology -- Periodicals; Molecular biology; Periodicals; 572.8
• DOI: 10.15252/embr.201541823
• Languages: English
• ISSNs: 1469-221X

Abstract

Abstract: Correct orientation of cell division is considered an important factor for the achievement of normal brain size, as mutations in genes that affect this process are among the leading causes of microcephaly. Abnormal spindle orientation is associated with reduction of the neuronal progenitor symmetric divisions, premature cell cycle exit, and reduced neurogenesis. This mechanism has been involved in microcephaly resulting from mutation of ASPM, the most frequently affected gene in autosomal recessive human primary microcephaly (MCPH), but it is presently unknown how ASPM regulates spindle orientation. In this report, we show that ASPM may control spindle positioning by interacting with citron kinase (CITK), a protein whose loss is also responsible for severe microcephaly in mammals. We show that the absence of CITK leads to abnormal spindle orientation in mammals and insects. In mouse cortical development, this phenotype correlates with increased production of basal progenitors. ASPM is required to recruit CITK at the spindle, and CITK overexpression rescues ASPM phenotype. ASPM and CITK affect the organization of astral microtubules (MT), and low doses of MT‐stabilizing drug revert the spindle orientation phenotype produced by their knockdown. Finally, CITK regulates both astral‐MT nucleation and stability. Our results provide a functional link between two established microcephaly proteins. Synopsis: The microcephaly gene ASPM recruits the cytokinesis regulator CITK to the mitotic spindle of neural progenitor cells, where it promotes the nucleation and stabilization of astral microtubules, providing a link between the two microcephaly proteins. The loss of CITK function in the developing mouse cortex results in increased cell cycle exit and generation of basal progenitors. CITK physically interacts with ASPM at the spindle pole in HeLa cells. CITK‐ and ASPM‐depleted HeLa cells display a significant reduction in the number and length of astral microtubules. Abstract : The microcephaly gene ASPM recruits the cytokinesis regulator CITK to the mitotic spindle of neural progenitor cells, where it promotes the nucleation and stabilization of astral microtubules, providing a link between the two microcephaly proteins.