Exportins can inhibit major mitotic assembly events in vitro: membrane fusion, nuclear pore formation, and spindle assembly

Autor:	Anna Travesa, Sarah Carmona, Dennis C. Garland, Cyril Bernis, Matthew S Nord, Douglass J. Forbes
Rok vydání:	2020
Předmět:	assembly lcsh:QH426-470 importins rangtp Mitosis Spindle Apparatus Importin Membrane Fusion environment and public health Xenopus laevis 03 medical and health sciences medicine Animals Humans lcsh:QH573-671 Nuclear pore Nuclear membrane crm1 030304 developmental biology karyopherins 0303 health sciences Cell-Free System lcsh:Cytology Chemistry fungi 030302 biochemistry & molecular biology Lipid bilayer fusion spindle nucleoporin Cell Biology Cell biology exportins lcsh:Genetics medicine.anatomical_structure nuclear pore Karyopherins Ran cell cycle Nucleoporin Research Article Research Paper
Zdroj:	Nucleus, Vol 11, Iss 1, Pp 178-193 (2020) Nucleus article-version (VoR) Version of Record
ISSN:	1949-1042 1949-1034
DOI:	10.1080/19491034.2020.1798093
Popis:	Xenopus egg extracts are a powerful in vitro tool for studying complex biological processes, including nuclear reconstitution, nuclear membrane and pore assembly, and spindle assembly. Extracts have been further used to demonstrate a moonlighting regulatory role for nuclear import receptors or importins on these cell cycle assembly events. Here we show that exportins can also play a role in these events. Addition of Crm1, Exportin-t, or Exportin-5 decreased nuclear pore assembly in vitro. RanQ69L-GTP, a constitutively active form of RanGTP, ameliorated inhibition. Both Crm1 and Exportin-t inhibited fusion of nuclear membranes, again counteracted by RanQ69L-GTP. In mitotic extracts, Crm1 and Exportin-t negatively impacted spindle assembly. Pulldowns from the extracts using Crm1- or Exportin-t-beads revealed nucleoporins known to be essential for both nuclear pore and spindle assembly, with RanQ69L-GTP decreasing a subset of these target interactions. This study suggests a model where exportins, like importins, can regulate major mitotic assembly events.
Databáze:	OpenAIRE
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