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

Autor: Matthew S. Nord, Cyril Bernis, Sarah Carmona, Dennis C. Garland, Anna Travesa, Douglass J. Forbes
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: Nucleus, Vol 11, Iss 1, Pp 178-193 (2020)
Druh dokumentu: article
ISSN: 1949-1034
1949-1042
19491034
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: Directory of Open Access Journals
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