A survey of the specificity and mechanism of 1,6 hexanediol-induced disruption of nuclear transport

ABSTRACT

Selective transport through the nuclear pore complex (NPC) depends on the dynamic binding of FG-repeat containing nucleoporins, the FG-nups, with each other and with Karyopherins (Kaps). Here, we assessed the specificity and mechanism by which the aliphatic alcohol 1,6-hexanediol (1,6HD) disrupts the permeability barrier of NPCs in live baker’s yeast cells. After a 10-minute exposure to 5% 1,6HD, no notable changes were observed in cell growth, cytosolic pH and ATP levels, or the appearance of organelles. However, effects on the cytoskeleton and Hsp104 were noted. 1,6HD clearly affected the NPC permeability barrier, allowing passive nuclear entry of a 177kDa reporter protein that is normally confined to the cytosol. Moreover, multiple Kaps were displaced from NPCs, and the displacement of Kap122-GFP correlated with the observed passive permeability changes. 1,6HD thus temporarily permeates NPCs, and in line with Kap-centric models, the mechanism includes the release of numerous Kaps from the NPCs.

KEYWORDS:

• 1,6-hexanediol
• aliphatic alcohol
• baker’s yeast
• Karyopherin
• liquid-liquid phase separation
• Nuclear pore complex
• nuclear transport
• Nuclear transport receptors

Acknowledgments

We want to thank Amarins Blaauwbroek for practical assistance.

Disclosure statement

No potential conflict of interest was reported by the author(s). The authors declare no competing interests.

Author contributions

ERB and LMV conceived the project. ERB designed, performed and analyzed all experiments with help from SNM (Figure 2B,C) and TO (Supplementary Figure S2). The manuscript was written by ERB and LMV with input of all authors.

Reagent availability

All reagents are available upon request.

SUPPLEMENTARY MATERIAL

Supplemental data for this article can be accessed online at https://doi.org/10.1080/19491034.2023.2240139

Additional information

Funding

ERB and TO are supported by PhD-fellowships from the Graduate School of Medical Sciences of the University of Groningen. ERB, AS, LMV, are supported by a Vici grant (VI.C.192.031) from the Netherlands Organisation for Scientific Research.