Loss of MYO5B expression deregulates late endosome size which hinders mitotic spindle orientation

Changsen Leng; Arend W Overeem; Fernando Cartón-Garcia; Qinghong Li; Karin Klappe; Jeroen Kuipers; Yingying Cui; Inge S Zuhorn; Diego Arango; Sven C D van IJzendoorn

doi:10.1371/journal.pbio.3000531

Loss of MYO5B expression deregulates late endosome size which hinders mitotic spindle orientation

PLoS Biol. 2019 Nov 4;17(11):e3000531. doi: 10.1371/journal.pbio.3000531. eCollection 2019 Nov.

Affiliations

¹ Department of Biomedical Sciences of Cells and Systems, section Molecular Cell Biology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
² Group of Biomedical Research in Digestive Tract Tumors, CIBBIM-Nanomedicine, Vall d'Hebron Research Institute (VHIR), Universitat Autònoma de Barcelona (UAB), Barcelona, Spain.
³ Department of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
⁴ Department of Biomedical Engineering, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.

Abstract

Recycling endosomes regulate plasma membrane recycling. Recently, recycling endosome-associated proteins have been implicated in the positioning and orientation of the mitotic spindle and cytokinesis. Loss of MYO5B, encoding the recycling endosome-associated myosin Vb, is associated with tumor development and tissue architecture defects in the gastrointestinal tract. Whether loss of MYO5B expression affects mitosis is not known. Here, we demonstrate that loss of MYO5B expression delayed cytokinesis, perturbed mitotic spindle orientation, led to the misorientation of the plane of cell division during the course of mitosis, and resulted in the delamination of epithelial cells. Remarkably, the effects on spindle orientation, but not cytokinesis, were a direct consequence of physical hindrance by giant late endosomes, which were formed in a chloride channel-sensitive manner concomitant with a redistribution of chloride channels from the cell periphery to late endosomes upon loss of MYO5B. Rab7 availability was identified as a limiting factor for the development of giant late endosomes. In accordance, increasing rab7 availability corrected mitotic spindle misorientation and cell delamination in cells lacking MYO5B expression. In conclusion, we identified a novel role for MYO5B in the regulation of late endosome size control and identify the inability to control late endosome size as an unexpected novel mechanism underlying defects in cell division orientation and epithelial architecture.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Caco-2 Cells
Cell Adhesion / physiology
Cell Division / physiology
Cell Line
Cell Membrane / metabolism
Cytokinesis / genetics
Cytokinesis / physiology
Endosomes / genetics
Endosomes / metabolism*
Epithelial Cells / metabolism
Female
HEK293 Cells
Humans
Mice
Mice, Inbred C57BL
Mice, Knockout
Mitosis / physiology
Myosin Heavy Chains / genetics
Myosin Heavy Chains / metabolism*
Myosin Type V / genetics
Myosin Type V / metabolism*
Spindle Apparatus / metabolism*
rab GTP-Binding Proteins / metabolism
rab7 GTP-Binding Proteins

Substances

MYO5B protein, human
Myo5B protein, mouse
rab7 GTP-Binding Proteins
rab7 GTP-binding proteins, human
rab7 GTP-binding proteins, mouse
Myosin Type V
Myosin Heavy Chains
rab GTP-Binding Proteins

Grants and funding

AD: Association for International Cancer Research (AICR13-0245), https://www.aicr.org AD: European Regional Development Fund (ERDF; PI16/00540 and AC15/00066), https://ec.europa.eu/regional_policy/en/funding/erdf AD: Spanish Association Against Cancer (AECC GCA15152966ARAN), https://www.uicc.org AD: the Instituto de Salud Carlos III, https://www.isciii.es/Paginas/Inicio.aspx IZ: The Dutch Research Council, Domain Applied and Engineering Sciences, https://www.nwo.nl/en JK: The Netherlands Organisation for Health Research and Development, 91111.006, https://www.zonmw.nl/en/ JK: The Dutch Research Council, 175-010-2009-023, https://www.nwo.nl/en CK: Chinese Scholarship Council, https://www.chinesescholarshipcouncil.com/ QL: Chinese Scholarship Council, https://www.chinesescholarshipcouncil.com/ YC: Chinese Scholarship Council, https://www.chinesescholarshipcouncil.com/. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.