| 2017 |
The FG repeat domain of Nup54 forms labile cross-β polymers (characterized by chemical footprinting), and mutations within the footprinted region block both polymerization and binding by the toxic PRn poly-dipeptide encoded by C9orf72 repeat expansion. PRn binding locks FG repeats in the polymerized state, inhibiting nuclear import/export. |
Chemical footprinting of FG domain polymers, site-directed mutagenesis blocking polymerization, in vitro 1,6-hexanediol treatment to melt polymers |
Proceedings of the National Academy of Sciences of the United States of America |
High |
28069952
|
| 2017 |
In C. elegans, channel nucleoporins NPP-4/Nup54 (along with NPP-1/Nup58 and NPP-11/Nup62) recruit PLK-1 to nuclear pore complexes prior to nuclear envelope breakdown (NEBD) via physical interaction between multiple Cdk1/PLK-1-primed Polo-docking sites and the PLK-1 Polo-box domain (PBD). This localization is required for efficient NEBD. |
Co-IP/pulldown of PLK-1 PBD with nucleoporins, genetic loss-of-function (RNAi) with NEBD phenotype, phosphorylation-site mutagenesis, live imaging |
Developmental cell |
High |
29065307
|
| 2015 |
Nup54 is the central subunit of the Nup62 channel complex, directly binding both Nup62 and Nup58 via coiled-coil segments. The structured domain of Nup58 allosterically couples with its neighboring disordered domain in interaction with Nup54's disordered domain and transport factor Kapβ1; multivalent Kapβ1 interactions with disordered domains of Nup58 stabilize the structured domain associated with Nup54, shifting equilibria from homo-oligomers to hetero-oligomers, providing a mechanism for NPC central channel constriction/dilation. |
Crystallography, analytical ultracentrifugation, equilibrium binding analysis, in vitro reconstitution of complexes |
Cell |
High |
26046439
|
| 2014 |
The Nup62 complex (Nup62, Nup54, Nup58) has a 1:1:1 stoichiometry in solution, with Nup54 central and directly binding both Nup62 and Nup58 via conserved coiled-coil segments. This stoichiometry is conserved in the homologous yeast Nsp1 complex. |
Gel filtration chromatography, analytical ultracentrifugation, in vitro reconstitution |
Molecular biology of the cell |
High |
24574455
|
| 2015 |
The ordered regions of Nup62, Nup54, and Nup58 form a dynamic 'triple complex' in solution built from characterized Nup54·Nup58 and Nup54·Nup62 interactomes. Nup54·Nup58 segments form inter-convertible hetero- and homo-oligomers proposed as 'mid-plane rings' for NPC channel dilation/constriction, while Nup54·Nup62 forms 1:2 triple-helix 'fingers' projecting from the ring. |
Solution analysis (SEC, AUC, SAXS), crystal structure analysis, in vitro reconstitution of sub-complexes |
The Journal of biological chemistry |
High |
26025361
|
| 2007 |
In Drosophila, RNAi-mediated inactivation of Nup54 (fly homolog) selectively impairs importin α/β-mediated nuclear import but not CRM1-dependent export, identifying Nup54 FG nucleoporin as specifically required for the import pathway. |
RNAi knockdown in Drosophila S2 cells, nuclear import/export reporter assays |
The Journal of cell biology |
Medium |
17682050
|
| 2018 |
Nup54 (along with its complex partners Nup62 and Nup58) is required for homologous recombination (HR) repair. Nup54 depletion decreased HR repair activity (measured by specific reporters), reduced HR-linked DNA synthesis foci and sister chromatid exchanges after ionizing radiation, and increased chromosome aberrations. Nup54 is epistatic with HR factor Rad51, and its loss caused mitotic catastrophe and enhanced radiosensitivity specifically in cells with replicated DNA. |
siRNA knockdown, HR reporter assays, FACS cell cycle analysis, chromosome aberration analysis, epistasis analysis with Rad51, ionizing radiation survival assays |
Nucleic acids research |
High |
29986057
|
| 2021 |
In Drosophila ovarian follicle cells, Nup54 (along with Nup58) is specifically required for piRNA biogenesis from the flamenco locus, with loss of Nup54/Nup58 causing compromised piRNA production exclusively from this locus while knockdown of other NPC subunits has more widespread consequences, indicating a specialized role beyond general nuclear transport. |
RNAi knockdown in Drosophila ovary, small RNA sequencing, piRNA pathway reporter assays |
eLife |
Medium |
33856346
|
| 2021 |
In Drosophila, Nup54 loss-of-function mutations cause defective wiring of eight adult brain pickpocket-expressing neurons required for egg-laying, and reduce sensitivity to sex-peptide, indicating a role for Nup54 in neuronal circuit differentiation underlying the female post-mating response. Nup54 promoter variants alter nucleo-cytoplasmic shuttling. |
Forward genetic screen, viable mutant allele characterization, neuroanatomical analysis (brain neuron wiring), behavioral assays (egg-laying, sex-peptide sensitivity) |
BMC biology |
Medium |
34666772
|
| 2021 |
Nup54 promotes nuclear importation of the arginine methyltransferase CARM1; Nup54 was identified as a CARM1-interacting protein, and overexpression of Nup54 facilitates CARM1 nuclear import, which in turn activates Notch2 transcription and methylation in gastric cancer cells. |
Co-immunoprecipitation identifying Nup54-CARM1 interaction, nuclear fractionation showing CARM1 localization changes, functional proliferation and tumor formation assays |
Oncogene |
Medium |
34725461
|
| 2017 |
Expression of Nup54 (or Nup62) rescues nuclear import of a TDP-43 NLS reporter that is impaired by cytoplasmic poly-GA aggregates in C9orf72 ALS/FTLD, placing Nup54 in the importin-α/β-dependent nuclear import pathway as a downstream effector. |
Rescue/overexpression experiments in cell culture with fluorescent NLS reporter, nuclear import quantification |
Human molecular genetics |
Medium |
28040728
|
| 2020 |
In live cells, conformational changes of Nup54 (along with Nup58 and Nup62) within the NPC inner ring are detected when transport through the NPC is perturbed, while Nups elsewhere in the NPC show no such changes, indicating that these inner ring nucleoporins are flexible and undergo transport-state-dependent conformational changes. |
FRET-based conformational sensors with rigidly conjugated mEGFP on NPC proteins in live cells, transport perturbation assays |
eLife |
Medium |
33346731
|
| 2022 |
TIP60-mediated acetylation of Nup62 at Lys432 dissolves the Nup62-Nup58-Nup54 complex during mitotic entry, promoting Nup62 redistribution to the mitotic spindle. This remodeling of the nucleoporin complex is required for correct spindle orientation and accurate chromosome segregation. |
Identification of TIP60 as acetyltransferase by biochemical assay, site-specific mutagenesis of Lys432, co-immunoprecipitation of Nup62-Nup58-Nup54 complex disruption, live imaging, spindle orientation and chromosome segregation assays |
Journal of molecular cell biology |
Medium |
36190325
|
| 2022 |
Biallelic (recessive) variants in NUP54 clustered in the C-terminal region that interacts with NUP62 cause early-onset dystonia with striatal lesions in humans, a phenotype similar to NUP62-related infantile striatonigral degeneration, establishing that the NUP54–NUP62 interaction is required for NPC function in the basal ganglia. |
Patient sequencing, in silico variant analysis, protein-biochemical studies confirming pathogenicity of variants affecting NUP62-interaction domain |
Annals of neurology |
Medium |
36333996
|
| 2011 |
NUP54 was identified as a host factor required for influenza virus polymerase replication/transcriptional activity in human cells, placing it in the nuclear transport function supporting viral RNA polymerase activity. |
Yeast two-hybrid screen for influenza polymerase interactors, siRNA knockdown validation of virus replication |
Journal of virology |
Low |
21994455
|
| 2023 |
In C. elegans, nuclear pore component NPP-1/Nup54 promotes perinuclear localization of the anti-silencing CSR-1 Argonaute and is required for piRNA-mediated gene silencing initiation, linking NPC function to piRNA pathway regulation. |
Sensitized piRNA reporter screen, RNAi knockdown, fluorescence microscopy of Argonaute localization |
Genetics |
Low |
37210214
|