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PreprintCited by 29Nuclear Structure and FunctionMicrotubule and mitosis dynamicsRNA Research and Splicing

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An amphipathic helix in Brl1 is required for membrane fusion during nuclear pore complex biogenesis in S. cerevisiae

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Kralt, Annemarie, Wojtynek, Matthias, Fischer, Jonas S., Agote-Aran, Arantxa, Mancini, Roberta, Dultz, Elisa, Noor, Elad, Uliana, Federico, Tatarek-Nossol, Marianna, Antonin, Wolfram, Onischenko, Evgeny, Medalia, Ohad, Weis, Karsten·2022·eLife
DOI 10.1101/2022.03.04.483005PMC PMC9402233

Preprint — published as 10.7554/elife.78385

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Abstract

The nuclear pore complex (NPC) is the central portal for macromolecular exchange between the nucleus and cytoplasm. In all eukaryotes, NPCs assemble into an intact nuclear envelope (NE) during interphase, but the process of NPC biogenesis remains poorly characterized. Furthermore, little is known about how NPC assembly leads to the fusion of the outer and inner NE, and no factors have been identified that could trigger this event. Here we characterize the transmembrane protein Brl1 as an NPC assembly factor required for NE fusion in budding yeast. Brl1 preferentially associates with NPC assembly intermediates and its depletion halts NPC biogenesis, leading to NE herniations that contain inner and outer ring nucleoporins but lack the cytoplasmic export platform. Furthermore, we identify an essential amphipathic helix in the luminal domain of Brl1 that mediates interactions with lipid bilayers. Mutations in this amphipathic helix lead to NPC assembly defects, and cryo-ET analyses reveal multi-layered herniations of the inner nuclear membrane with NPC-like structures at the neck, indicating a failure in NE fusion. Taken together, our results identify a role for Brl1 in NPC assembly and suggest a function of its amphipathic helix in mediating the fusion of the inner and outer nuclear membranes.

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Metadata

Authors
Kralt, A., Wojtynek, M., Fischer, J. S., Agote-Aran, A., Mancini, R., Dultz, E., Noor, E., Uliana, F., Tatarek-Nossol, M., Antonin, W., Onischenko, E., Medalia, O., & Weis, K.
Affiliations
  • ETH Zurich (CH)
  • University of Zurich (CH)
  • RWTH Aachen University (DE)
  • University of Bergen (NO)
Corresponding author
Weis, Karsten
ORCID
Venue
eLife
Publisher
openRxiv
Published
2022-03-04
Volume
11
ISSN
2050-084X (ISSN-L)
DOI
10.1101/2022.03.04.483005
PMC
PMC9402233
OpenAlex
W4293101369
Cited by
29
References
1
Citations by year
2026: 10 · 2025: 5 · 2024: 4 · 2023: 6 · 2022: 4
FWCI
1.55
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • eLife (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Zurich Open Repository and Archive (University of Zurich) (acceptedVersion, cc-by)
  • NORA - Norwegian Open Research Archives (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • RWTH Publications (RWTH Aachen) (submittedVersion, other-oa)
  • Repository for Publications and Research Data (ETH Zurich) (submittedVersion, cc-by)
  • Zurich Open Repository and Archive (University of Zurich) (submittedVersion, cc-by)
  • RWTH Publications (RWTH Aachen)
  • ETH Zürich Research Collection
  • Zurich Open Repository and Archive (University of Zurich)
Funders
  • National Science Foundation
  • Deutsche Forschungsgemeinschaft (AN377/7‐1, DFG AN377/7-1)
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (171317, 179275, 179418)
  • Eidgenössische Technische Hochschule Zürich
  • Norges Forskningsråd (NFR 315615)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Keywords
amphipathic helix, nucleoporins, lipid bilayer binding, transmembrane proteins, nuclear envelope assembly, Saccharomyces cerevisiae
MeSH terms
Nuclear Envelope, Saccharomyces cerevisiae, Nuclear Pore, Nuclear Pore Complex Proteins, Saccharomyces cerevisiae Proteins

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