Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics
Phosphorylation of the C-terminal tail of the heavy neurofilament subunit (NF-H) impacts neurofilament (NF) axonal transport and residence within axons by fostering NF-NF associations that compete with transport. We tested the role of phosphorylation of a GSK-3β consensus site (S493) located in the...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2017-10-01
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| Series: | Biology Open |
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| Online Access: | http://bio.biologists.org/content/6/10/1516 |
| _version_ | 1818865603057811456 |
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| author | Rishel Brenna Vohnoutka Edward F. Boumil Yuguan Liu Atsuko Uchida Harish C. Pant Thomas B. Shea |
| author_facet | Rishel Brenna Vohnoutka Edward F. Boumil Yuguan Liu Atsuko Uchida Harish C. Pant Thomas B. Shea |
| author_sort | Rishel Brenna Vohnoutka |
| collection | DOAJ |
| description | Phosphorylation of the C-terminal tail of the heavy neurofilament subunit (NF-H) impacts neurofilament (NF) axonal transport and residence within axons by fostering NF-NF associations that compete with transport. We tested the role of phosphorylation of a GSK-3β consensus site (S493) located in the proximal portion of the NF-H tail in NF dynamics by transfection of NB2a/d1 cells with NF-H, where S493 was mutated to aspartic acid (S493D) or to alanine (S493A) to mimic constitutive phosphorylation and non-phosphorylation. S493D underwent increased transport into axonal neurites, while S493A displayed increased perikaryal NF aggregates that were decorated by anti-kinesin. Increased levels of S493A co-precipitated with anti-kinesin indicating that reduced transport of S493A was not due to reduced kinesin association but due to premature NF-NF interactions within perikarya. S493D displayed increased phospho-immunoreactivity within axonal neurites at downstream C-terminal sites attributable to mitogen-activated protein kinase and cyclin-dependent kinase 5. However, S493D was more prone to proteolysis following kinase inhibition, suggesting that S493 phosphorylation is an early event that alters sidearm configuration in a manner that promotes appropriate NF distribution. We propose a novel model for sidearm configuration. |
| first_indexed | 2024-12-19T10:50:10Z |
| format | Article |
| id | doaj.art-e0abb5973e3f4fbbbda52cbc17595979 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-19T10:50:10Z |
| publishDate | 2017-10-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-e0abb5973e3f4fbbbda52cbc175959792022-12-21T20:25:03ZengThe Company of BiologistsBiology Open2046-63902017-10-016101516152710.1242/bio.028522028522Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamicsRishel Brenna Vohnoutka0Edward F. Boumil1Yuguan Liu2Atsuko Uchida3Harish C. Pant4Thomas B. Shea5 Laboratory for Neuroscience, University of Massachusetts Lowell, Lowell, MA 01854, USA Laboratory for Neuroscience, University of Massachusetts Lowell, Lowell, MA 01854, USA Department of Biomedical and Nutritional Sciences, University of Massachusetts Lowell, Lowell, MA 01854, USA Department of Neuroscience, Ohio State University, Columbus, OH 43210, USA Cytoskeletal Protein Regulation Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA Laboratory for Neuroscience, University of Massachusetts Lowell, Lowell, MA 01854, USA Phosphorylation of the C-terminal tail of the heavy neurofilament subunit (NF-H) impacts neurofilament (NF) axonal transport and residence within axons by fostering NF-NF associations that compete with transport. We tested the role of phosphorylation of a GSK-3β consensus site (S493) located in the proximal portion of the NF-H tail in NF dynamics by transfection of NB2a/d1 cells with NF-H, where S493 was mutated to aspartic acid (S493D) or to alanine (S493A) to mimic constitutive phosphorylation and non-phosphorylation. S493D underwent increased transport into axonal neurites, while S493A displayed increased perikaryal NF aggregates that were decorated by anti-kinesin. Increased levels of S493A co-precipitated with anti-kinesin indicating that reduced transport of S493A was not due to reduced kinesin association but due to premature NF-NF interactions within perikarya. S493D displayed increased phospho-immunoreactivity within axonal neurites at downstream C-terminal sites attributable to mitogen-activated protein kinase and cyclin-dependent kinase 5. However, S493D was more prone to proteolysis following kinase inhibition, suggesting that S493 phosphorylation is an early event that alters sidearm configuration in a manner that promotes appropriate NF distribution. We propose a novel model for sidearm configuration.http://bio.biologists.org/content/6/10/1516Axonal cytoskeletonNeurofilamentPhosphorylationProtein conformationProteolysis |
| spellingShingle | Rishel Brenna Vohnoutka Edward F. Boumil Yuguan Liu Atsuko Uchida Harish C. Pant Thomas B. Shea Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics Biology Open Axonal cytoskeleton Neurofilament Phosphorylation Protein conformation Proteolysis |
| title | Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics |
| title_full | Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics |
| title_fullStr | Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics |
| title_full_unstemmed | Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics |
| title_short | Influence of a GSK3β phosphorylation site within the proximal C-terminus of Neurofilament-H on neurofilament dynamics |
| title_sort | influence of a gsk3β phosphorylation site within the proximal c terminus of neurofilament h on neurofilament dynamics |
| topic | Axonal cytoskeleton Neurofilament Phosphorylation Protein conformation Proteolysis |
| url | http://bio.biologists.org/content/6/10/1516 |
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