Nuclear export of the pre-60S ribosomal subunit through single nuclear pores observed in real time

Jan Andreas Ruland, Annika Marie Krüger, Kerstin Dörner, Rohan Bhatia, Sabine Wirths, Daniel Poetes, Ulrike Kutay, Jan Peter Siebrasse and Ulrich Kubitscheck ()
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Jan Andreas Ruland: University of Bonn
Annika Marie Krüger: University of Bonn
Kerstin Dörner: ETH Zurich
Rohan Bhatia: University of Bonn
Sabine Wirths: University of Bonn
Daniel Poetes: University of Bonn
Ulrike Kutay: ETH Zurich
Jan Peter Siebrasse: University of Bonn
Ulrich Kubitscheck: University of Bonn

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract Ribosomal biogenesis has been studied by biochemical, genetic and electron microscopic approaches, but live cell data on the in vivo kinetics are still missing. Here we analyse the export kinetics of the large ribosomal subunit (pre-60S particle) through single NPCs in human cells. We established a stable cell line co-expressing Halo-tagged eIF6 and GFP-fused NTF2 to simultaneously label pre-60S particles and NPCs, respectively. By combining single molecule tracking and super resolution confocal microscopy we visualize the dynamics of single pre-60S particles during export through single NPCs. For export events, maximum particle accumulation is found in the centre of the pore, while unsuccessful export terminates within the nuclear basket. The export has a single rate limiting step and a duration of ∼24 milliseconds. Only about 1/3 of attempted export events are successful. Our results show that the mass flux through a single NPC can reach up to ~125 MDa·s−1 in vivo.

Date: 2021
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-26323-7

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DOI: 10.1038/s41467-021-26323-7

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