Importin-β negatively regulates multiple aspects of mitosis including RANGAP1 recruitment to kinetochores.

The Journal of Cell Biology
Emanuele RoscioliPatrizia Lavia

Abstract

Importin-β is the main vector for interphase nuclear protein import and plays roles after nuclear envelope breakdown. Here we show that importin-β regulates multiple aspects of mitosis via distinct domains that interact with different classes of proteins in human cells. The C-terminal region (which binds importin-α) inhibits mitotic spindle pole formation. The central region (harboring nucleoporin-binding sites) regulates microtubule dynamic functions and interaction with kinetochores. Importin-β interacts through this region with NUP358/RANBP2, which in turn binds SUMO-conjugated RANGAP1 in nuclear pores. We show that this interaction continues after nuclear pore disassembly. Overexpression of importin-β, or of the nucleoporin-binding region, inhibited RANGAP1 recruitment to mitotic kinetochores, an event that is known to require microtubule attachment and the exportin CRM1. Co-expressing either importin-β-interacting RANBP2 fragments, or CRM1, restored RANGAP1 to kinetochores and rescued importin-β-dependent mitotic dynamic defects. These results reveal previously unrecognized importin-β functions at kinetochores exerted via RANBP2 and opposed by CRM1.

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Citations

Aug 31, 2013·Neuromolecular Medicine·Mathias DroescherAndrea Pichler
Jan 5, 2016·Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology·Patrizia Lavia
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Jun 11, 2017·Journal of Cell Science·Eugenia GilistroPatrizia Lavia
Nov 27, 2018·ELife·Claudia PellacaniMaria Patrizia Somma
Jul 7, 2020·Frontiers in Cell and Developmental Biology·Ly T S Nguyen, Douglas N Robinson
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Aug 14, 2021·Journal of Cell Science·Takanobu Moriuchi, Fumiko Hirose

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Methods Mentioned

BETA
GTPase
transfection
coIP
FACS
PCR

Software Mentioned

NIS
ISO
Mascot Search engine
ACT
Elements
Elements AR
Mascot
PeakErazor
ImageJ
CREST

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