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Entry into the nuclear pore complex is controlled by a cytoplasmic exclusion zone containing dynamic GLFG-repeat nucleoporin domains

Fiserova, J.; Spink, M.; Richards, S.A.; Saunter, C.; Goldberg, M.W.

Entry into the nuclear pore complex is controlled by a cytoplasmic exclusion zone containing dynamic GLFG-repeat nucleoporin domains Thumbnail


Authors

J. Fiserova

M. Spink

S.A. Richards

C. Saunter



Abstract

Nuclear pore complexes (NPCs) mediate nucleocytoplasmic movement. The central channel contains proteins with phenylalanine-glycine (FG) repeats, or variations (GLFG, glycine-leucine-phenylalanine-glycine). These are ‘intrinsically disordered’ and often represent weak interaction sites that become ordered upon interaction. We investigated this possibility during nuclear transport. Using electron microscopy of S. cerevisiae, we show that NPC cytoplasmic filaments form a dome-shaped structure enclosing GLFG domains. GLFG domains extend out of this structure and are part of an ‘exclusion zone’ that might act as a partial barrier to entry of transport-inert proteins. The anchor domain of a GLFG nucleoporin locates exclusively to the central channel. By contrast, the localisation of the GLFG domains varied between NPCs and could be cytoplasmic, central or nucleoplasmic and could stretch up to 80 nm. These results suggest a dynamic exchange between ordered and disordered states. In contrast to diffusion through the NPC, transport cargoes passed through the exclusion zone and accumulated near the central plane. We also show that movement of cargo through the NPC is accompanied by relocation of GLFG domains, suggesting that binding, restructuring and movement of these domains could be part of the translocation mechanism.

Citation

Fiserova, J., Spink, M., Richards, S., Saunter, C., & Goldberg, M. (2014). Entry into the nuclear pore complex is controlled by a cytoplasmic exclusion zone containing dynamic GLFG-repeat nucleoporin domains. Journal of Cell Science, 127, 124-136. https://doi.org/10.1242/jcs.133272

Journal Article Type Article
Acceptance Date Sep 29, 2013
Online Publication Date Jan 1, 2014
Publication Date Jan 1, 2014
Deposit Date Oct 1, 2013
Publicly Available Date Aug 22, 2014
Journal Journal of Cell Science
Print ISSN 0021-9533
Electronic ISSN 1477-9137
Publisher The Company of Biologists
Peer Reviewed Peer Reviewed
Volume 127
Pages 124-136
DOI https://doi.org/10.1242/jcs.133272
Keywords Nuclear pore, Transport, Phenylalanine-glycine repeat, GLFG.

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