title: Automated tracking of mitotic spindle pole positions shows that LGN is required for spindle rotation but not orientation maintenance.
abstract: Spindle orientation defines the plane of cell division and, thereby, the spatial position of all daughter cells. Here, we develop a live cell microscopy-based methodology to extract spindle movements in human epithelial cell lines and study how spindles are brought to a pre-defined orientation. We show that spindles undergo two distinct regimes of movements. Spindles are first actively rotated toward the cells' long-axis and then maintained along this pre-defined axis. By quantifying spindle movements in cells depleted of LGN, we show that the first regime of rotational movements requires LGN that recruits cortical dynein. In contrast, the second regime of movements that maintains spindle orientation does not require LGN, but is sensitive to 2ME2 that suppresses microtubule dynamics. Our study sheds first insight into spatially defined spindle movement regimes in human cells, and supports the presence of LGN and dynein independent cortical anchors for astral microtubules.
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: Adam M
authors.initials: AM
authors.lastname: Corrigan
authors.forename: Roshan L
authors.initials: RL
authors.lastname: Shrestha
authors.forename: Ihsan
authors.initials: I
authors.lastname: Zulkipli
authors.forename: Noriko
authors.initials: N
authors.lastname: Hiroi
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Liu
authors.forename: Naoka
authors.initials: N
authors.lastname: Tamura
authors.forename: Bing
authors.initials: B
authors.lastname: Yang
authors.forename: Jessica
authors.initials: J
authors.lastname: Patel
authors.forename: Akira
authors.initials: A
authors.lastname: Funahashi
authors.forename: Athene
authors.initials: A
authors.lastname: Donald
authors.forename: Viji M
authors.initials: VM
authors.lastname: Draviam
mesh_terms: D004847:Epithelial Cells / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015151:Immunoblotting; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D018715:Microscopy, Video / Q000379:methods; D034741:RNA, Small Interfering / Q000235:genetics; D012399:Rotation; D008941:Spindle Apparatus / Q000502:physiology*
chemical_list: C102754:GPSM2 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D034741:RNA, Small Interfering
keywords: microtubule; mitosis; spindle orientation
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