bims-nucpor 2024-04-28 papers

Issue of 2024‒04‒28
four papers selected by
Sara Mingu, Johannes Gutenberg University

EMBO J. 2024 Apr 22.

A checkpoint function for Nup98 in nuclear pore formation suggested by novel inhibitory nanobodies.

Mireia Solà Colom, Zhenglin Fu, Philip Gunkel, Thomas Güttler, Sergei Trakhanov, Vasundara Srinivasan, Kathrin Gregor, Tino Pleiner, Dirk Görlich.

Nuclear pore complex (NPC) biogenesis is a still enigmatic example of protein self-assembly. We now introduce several cross-reacting anti-Nup nanobodies for imaging intact nuclear pore complexes from frog to human. We also report a simplified assay that directly tracks postmitotic NPC assembly with added fluorophore-labeled anti-Nup nanobodies. During interphase, NPCs are inserted into a pre-existing nuclear envelope. Monitoring this process is challenging because newly assembled NPCs are indistinguishable from pre-existing ones. We overcame this problem by inserting Xenopus-derived NPCs into human nuclear envelopes and using frog-specific anti-Nup nanobodies for detection. We further asked whether anti-Nup nanobodies could serve as NPC assembly inhibitors. Using a selection strategy against conserved epitopes, we obtained anti-Nup93, Nup98, and Nup155 nanobodies that block Nup-Nup interfaces and arrest NPC assembly. We solved structures of nanobody-target complexes and identified roles for the Nup93 α-solenoid domain in recruiting Nup358 and the Nup214·88·62 complex, as well as for Nup155 and the Nup98 autoproteolytic domain in NPC scaffold assembly. The latter suggests a checkpoint linking pore formation to the assembly of the Nup98-dominated permeability barrier.

Keywords: FG Phase; Nuclear Envelope; Nuclear Pores; Nuclear Transport; VHH Antibodies

DOI: https://doi.org/10.1038/s44318-024-00081-w

J Cell Biol. 2024 Jul 01. pii: e202310006. [Epub ahead of print]223(7):

UBAP2L ensures homeostasis of nuclear pore complexes at the intact nuclear envelope.

Yongrong Liao, Leonid Andronov, Xiaotian Liu, Junyan Lin, Lucile Guerber, Linjie Lu, Arantxa Agote-Arán, Evanthia Pangou, Li Ran, Charlotte Kleiss, Mengdi Qu, Stephane Schmucker, Luca Cirillo, Zhirong Zhang, Daniel Riveline, Monica Gotta, Bruno P Klaholz, Izabela Sumara.

Assembly of macromolecular complexes at correct cellular sites is crucial for cell function. Nuclear pore complexes (NPCs) are large cylindrical assemblies with eightfold rotational symmetry, built through hierarchical binding of nucleoporins (Nups) forming distinct subcomplexes. Here, we uncover a role of ubiquitin-associated protein 2-like (UBAP2L) in the assembly and stability of properly organized and functional NPCs at the intact nuclear envelope (NE) in human cells. UBAP2L localizes to the nuclear pores and facilitates the formation of the Y-complex, an essential scaffold component of the NPC, and its localization to the NE. UBAP2L promotes the interaction of the Y-complex with POM121 and Nup153, the critical upstream factors in a well-defined sequential order of Nups assembly onto NE during interphase. Timely localization of the cytoplasmic Nup transport factor fragile X-related protein 1 (FXR1) to the NE and its interaction with the Y-complex are likewise dependent on UBAP2L. Thus, this NPC biogenesis mechanism integrates the cytoplasmic and the nuclear NPC assembly signals and ensures efficient nuclear transport, adaptation to nutrient stress, and cellular proliferative capacity, highlighting the importance of NPC homeostasis at the intact NE.

DOI: https://doi.org/10.1083/jcb.202310006

FASEB J. 2024 Apr 30. 38(8): e23623

Karyopherin α2 is a maternal effect gene required for early embryonic development and female fertility in mice.

Franziska Rother, Reinhard Depping, Elena Popova, Stefanie Huegel, Ariane Heiler, Enno Hartmann, Michael Bader.

The nuclear transport of proteins plays an important role in mediating the transition from egg to embryo and distinct karyopherins have been implicated in this process. Here, we studied the impact of KPNA2 deficiency on preimplantation embryo development in mice. Loss of KPNA2 results in complete arrest at the 2cell stage and embryos exhibit the inability to activate their embryonic genome as well as a severely disturbed nuclear translocation of Nucleoplasmin 2. Our findings define KPNA2 as a new maternal effect gene.

Keywords: female fertility; importin; karyopherin; preimplantation development

DOI: https://doi.org/10.1096/fj.202301572RR

Acta Neuropathol Commun. 2024 Apr 25. 12(1): 69

LINC complex alterations are a key feature of sporadic and familial ALS/FTD.

Riccardo Sirtori, Michelle J Gregoire, Emily M Potts, Alicia Collins, Liviana Donatelli, Claudia Fallini.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder that primarily affects motor neurons, leading to progressive muscle weakness and loss of voluntary muscle control. While the exact cause of ALS is not fully understood, emerging research suggests that dysfunction of the nuclear envelope (NE) may contribute to disease pathogenesis and progression. The NE plays a role in ALS through several mechanisms, including nuclear pore defects, nucleocytoplasmic transport impairment, accumulation of mislocalized proteins, and nuclear morphology abnormalities. The LINC complex is the second biggest multi-protein complex in the NE and consists of the SUN1/2 proteins spanning the inner nuclear membrane and Nesprin proteins embedded in the outer membrane. The LINC complex, by interacting with both the nuclear lamina and the cytoskeleton, transmits mechanical forces to the nucleus regulating its morphology and functional homeostasis. In this study we show extensive alterations to the LINC complex in motor and cortical iPSC-derived neurons and spinal cord organoids carrying the ALS causative mutation in the C9ORF72 gene (C9). Importantly, we show that such alterations are present in vivo in a cohort of sporadic ALS and C9-ALS postmortem spinal cord and motor cortex specimens. We also found that LINC complex disruption strongly correlated with nuclear morphological alterations occurring in ALS neurons, independently of TDP43 mislocalization. Altogether, our data establish morphological and functional alterations to the LINC complex as important events in ALS pathogenic cascade, making this pathway a possible target for both biomarker and therapy development.

Keywords: ALS; C9ORF72; FTD; LINC complex; Nesprin; SUN

DOI: https://doi.org/10.1186/s40478-024-01778-z