Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy
Abstract Background Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. W...
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-12-01
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| Series: | Molecular Neurodegeneration |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13024-022-00585-1 |
| _version_ | 1811204152034852864 |
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| author | Bilal Khalil Deepak Chhangani Melissa C. Wren Courtney L. Smith Jannifer H. Lee Xingli Li Christian Puttinger Chih-Wei Tsai Gael Fortin Dmytro Morderer Junli Gao Feilin Liu Chun Kim Lim Jingjiao Chen Ching-Chieh Chou Cara L. Croft Amanda M. Gleixner Christopher J. Donnelly Todd E. Golde Leonard Petrucelli Björn Oskarsson Dennis W. Dickson Ke Zhang James Shorter Shige H. Yoshimura Sami J. Barmada Diego E. Rincon-Limas Wilfried Rossoll |
| author_facet | Bilal Khalil Deepak Chhangani Melissa C. Wren Courtney L. Smith Jannifer H. Lee Xingli Li Christian Puttinger Chih-Wei Tsai Gael Fortin Dmytro Morderer Junli Gao Feilin Liu Chun Kim Lim Jingjiao Chen Ching-Chieh Chou Cara L. Croft Amanda M. Gleixner Christopher J. Donnelly Todd E. Golde Leonard Petrucelli Björn Oskarsson Dennis W. Dickson Ke Zhang James Shorter Shige H. Yoshimura Sami J. Barmada Diego E. Rincon-Limas Wilfried Rossoll |
| author_sort | Bilal Khalil |
| collection | DOAJ |
| description | Abstract Background Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. While TDP-43 proteinopathy has been associated with defects in nucleocytoplasmic transport, this process is still poorly understood. Here we study the role of karyopherin-β1 (KPNB1) and other nuclear import receptors in regulating TDP-43 pathology. Methods We used immunostaining, immunoprecipitation, biochemical and toxicity assays in cell lines, primary neuron and organotypic mouse brain slice cultures, to determine the impact of KPNB1 on the solubility, localization, and toxicity of pathological TDP-43 constructs. Postmortem patient brain and spinal cord tissue was stained to assess KPNB1 colocalization with TDP-43 inclusions. Turbidity assays were employed to study the dissolution and prevention of aggregation of recombinant TDP-43 fibrils in vitro. Fly models of TDP-43 proteinopathy were used to determine the effect of KPNB1 on their neurodegenerative phenotype in vivo. Results We discovered that several members of the nuclear import receptor protein family can reduce the formation of pathological TDP-43 aggregates. Using KPNB1 as a model, we found that its activity depends on the prion-like C-terminal region of TDP-43, which mediates the co-aggregation with phenylalanine and glycine-rich nucleoporins (FG-Nups) such as Nup62. KPNB1 is recruited into these co-aggregates where it acts as a molecular chaperone that reverses aberrant phase transition of Nup62 and TDP-43. These findings are supported by the discovery that Nup62 and KPNB1 are also sequestered into pathological TDP-43 aggregates in ALS/FTD postmortem CNS tissue, and by the identification of the fly ortholog of KPNB1 as a strong protective modifier in Drosophila models of TDP-43 proteinopathy. Our results show that KPNB1 can rescue all hallmarks of TDP-43 pathology, by restoring its solubility and nuclear localization, and reducing neurodegeneration in cellular and animal models of ALS/FTD. Conclusion Our findings suggest a novel NLS-independent mechanism where, analogous to its canonical role in dissolving the diffusion barrier formed by FG-Nups in the nuclear pore, KPNB1 is recruited into TDP-43/FG-Nup co-aggregates present in TDP-43 proteinopathies and therapeutically reverses their deleterious phase transition and mislocalization, mitigating neurodegeneration. Graphical Abstract |
| first_indexed | 2024-04-12T03:07:33Z |
| format | Article |
| id | doaj.art-bbd064334e3c44abbee70cb5d84f3100 |
| institution | Directory Open Access Journal |
| issn | 1750-1326 |
| language | English |
| last_indexed | 2024-04-12T03:07:33Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Molecular Neurodegeneration |
| spelling | doaj.art-bbd064334e3c44abbee70cb5d84f31002022-12-22T03:50:28ZengBMCMolecular Neurodegeneration1750-13262022-12-0117112710.1186/s13024-022-00585-1Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathyBilal Khalil0Deepak Chhangani1Melissa C. Wren2Courtney L. Smith3Jannifer H. Lee4Xingli Li5Christian Puttinger6Chih-Wei Tsai7Gael Fortin8Dmytro Morderer9Junli Gao10Feilin Liu11Chun Kim Lim12Jingjiao Chen13Ching-Chieh Chou14Cara L. Croft15Amanda M. Gleixner16Christopher J. Donnelly17Todd E. Golde18Leonard Petrucelli19Björn Oskarsson20Dennis W. Dickson21Ke Zhang22James Shorter23Shige H. Yoshimura24Sami J. Barmada25Diego E. Rincon-Limas26Wilfried Rossoll27Department of Neuroscience, Mayo ClinicDepartment of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of FloridaDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neurology, University of MichiganDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicGraduate School of Biostudies, Kyoto UniversityDepartment of Neuroscience, Mayo ClinicDepartment of Biology, Stanford UniversityDepartment of Neuroscience, Center for Translational Research in Neurodegenerative Disease, University of FloridaDepartment of Neurobiology, University of Pittsburgh School of MedicineDepartment of Neurobiology, University of Pittsburgh School of MedicineDepartment of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of FloridaDepartment of Neuroscience, Mayo ClinicDepartment of Neurology, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Neuroscience, Mayo ClinicDepartment of Biochemistry and Biophysics, Perelman School of Medicine, University of PennsylvaniaGraduate School of Biostudies, Kyoto UniversityDepartment of Neurology, University of MichiganDepartment of Neurology, McKnight Brain Institute, Norman Fixel Institute for Neurological Diseases, University of FloridaDepartment of Neuroscience, Mayo ClinicAbstract Background Cytoplasmic mislocalization and aggregation of TAR DNA-binding protein-43 (TDP-43) is a hallmark of the amyotrophic lateral sclerosis and frontotemporal dementia (ALS/FTD) disease spectrum, causing both nuclear loss-of-function and cytoplasmic toxic gain-of-function phenotypes. While TDP-43 proteinopathy has been associated with defects in nucleocytoplasmic transport, this process is still poorly understood. Here we study the role of karyopherin-β1 (KPNB1) and other nuclear import receptors in regulating TDP-43 pathology. Methods We used immunostaining, immunoprecipitation, biochemical and toxicity assays in cell lines, primary neuron and organotypic mouse brain slice cultures, to determine the impact of KPNB1 on the solubility, localization, and toxicity of pathological TDP-43 constructs. Postmortem patient brain and spinal cord tissue was stained to assess KPNB1 colocalization with TDP-43 inclusions. Turbidity assays were employed to study the dissolution and prevention of aggregation of recombinant TDP-43 fibrils in vitro. Fly models of TDP-43 proteinopathy were used to determine the effect of KPNB1 on their neurodegenerative phenotype in vivo. Results We discovered that several members of the nuclear import receptor protein family can reduce the formation of pathological TDP-43 aggregates. Using KPNB1 as a model, we found that its activity depends on the prion-like C-terminal region of TDP-43, which mediates the co-aggregation with phenylalanine and glycine-rich nucleoporins (FG-Nups) such as Nup62. KPNB1 is recruited into these co-aggregates where it acts as a molecular chaperone that reverses aberrant phase transition of Nup62 and TDP-43. These findings are supported by the discovery that Nup62 and KPNB1 are also sequestered into pathological TDP-43 aggregates in ALS/FTD postmortem CNS tissue, and by the identification of the fly ortholog of KPNB1 as a strong protective modifier in Drosophila models of TDP-43 proteinopathy. Our results show that KPNB1 can rescue all hallmarks of TDP-43 pathology, by restoring its solubility and nuclear localization, and reducing neurodegeneration in cellular and animal models of ALS/FTD. Conclusion Our findings suggest a novel NLS-independent mechanism where, analogous to its canonical role in dissolving the diffusion barrier formed by FG-Nups in the nuclear pore, KPNB1 is recruited into TDP-43/FG-Nup co-aggregates present in TDP-43 proteinopathies and therapeutically reverses their deleterious phase transition and mislocalization, mitigating neurodegeneration. Graphical Abstracthttps://doi.org/10.1186/s13024-022-00585-1Amyotrophic lateral sclerosisFrontotemporal dementiaTDP-43Nucleocytoplasmic transportImportinNuclear pore |
| spellingShingle | Bilal Khalil Deepak Chhangani Melissa C. Wren Courtney L. Smith Jannifer H. Lee Xingli Li Christian Puttinger Chih-Wei Tsai Gael Fortin Dmytro Morderer Junli Gao Feilin Liu Chun Kim Lim Jingjiao Chen Ching-Chieh Chou Cara L. Croft Amanda M. Gleixner Christopher J. Donnelly Todd E. Golde Leonard Petrucelli Björn Oskarsson Dennis W. Dickson Ke Zhang James Shorter Shige H. Yoshimura Sami J. Barmada Diego E. Rincon-Limas Wilfried Rossoll Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy Molecular Neurodegeneration Amyotrophic lateral sclerosis Frontotemporal dementia TDP-43 Nucleocytoplasmic transport Importin Nuclear pore |
| title | Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy |
| title_full | Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy |
| title_fullStr | Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy |
| title_full_unstemmed | Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy |
| title_short | Nuclear import receptors are recruited by FG-nucleoporins to rescue hallmarks of TDP-43 proteinopathy |
| title_sort | nuclear import receptors are recruited by fg nucleoporins to rescue hallmarks of tdp 43 proteinopathy |
| topic | Amyotrophic lateral sclerosis Frontotemporal dementia TDP-43 Nucleocytoplasmic transport Importin Nuclear pore |
| url | https://doi.org/10.1186/s13024-022-00585-1 |
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