Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface
Cofilin-actin rods are a well-documented stress response in neuronal cells and their persistence is frequently associated with neurodegenerative disease. However, the role of specific actin residues in promoting the formation of cofilin-actin rods and other anomalous cytoskeletal structures is large...
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Frontiers Media S.A.
2025-06-01
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| Series: | Frontiers in Cellular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fncel.2025.1543199/full |
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| author | Noah Mann Keerthana Surabhi Josephine Sharp Mary Phipps Maelee Becton Jahiem Hill Davis Roberts Erzsebet M. Szatmari Robert M. Hughes |
| author_facet | Noah Mann Keerthana Surabhi Josephine Sharp Mary Phipps Maelee Becton Jahiem Hill Davis Roberts Erzsebet M. Szatmari Robert M. Hughes |
| author_sort | Noah Mann |
| collection | DOAJ |
| description | Cofilin-actin rods are a well-documented stress response in neuronal cells and their persistence is frequently associated with neurodegenerative disease. However, the role of specific actin residues in promoting the formation of cofilin-actin rods and other anomalous cytoskeletal structures is largely unknown. As it is increasingly suspected that specific mutations and post-translation modifications of actin may promote neurodegenerative disease, characterizing the role of these residues in cytoskeletal dysregulation is highly relevant. In this study, we focus on the actin-ATP interface, which has been proposed as a key mediator of cofilin-actin rod formation and the propensity of actin to respond to cellular stress. Using a light and stress-gated reporter of cofilin-actin cluster formation, we determine the impact of mutants associated with Actin-ATP binding on the propensity of actin to form anomalous structures in the presence and absence of applied cellular stress. This study identifies actin mutants that promote anomalous actin inclusions in HeLa cells and characterizes the manifestation of these phenotypes in cortical neurons. Mutations to the ATP phosphate tail-binding region of actin (K18A, D154A, G158L, K213A) were found to be particularly disruptive to actin phenotypes, and in several instances promote disease-associated actin-rich structures such as cofilin-actin rods and Hirano bodies. We find that these mutant phenotypes are largely consistent between cell types and display highly unusual inclusions in cultured cortical neurons, without leading to nuclear fragmentation and apoptotic death of the transfected cells. These mutants strengthen the association of residue-specific changes in actin with large-scale phenotypic and functional changes in the cytoskeleton, further implicating them in neurodegenerative disease progression. |
| format | Article |
| id | doaj-art-a918050d3cf641c2bbc0c944bc355b36 |
| institution | DOAJ |
| issn | 1662-5102 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj-art-a918050d3cf641c2bbc0c944bc355b362025-08-20T03:07:21ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022025-06-011910.3389/fncel.2025.15431991543199Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interfaceNoah Mann0Keerthana Surabhi1Josephine Sharp2Mary Phipps3Maelee Becton4Jahiem Hill5Davis Roberts6Erzsebet M. Szatmari7Robert M. Hughes8Department of Chemistry, East Carolina University, Greenville, NC, United StatesDepartment of Physical Therapy, East Carolina University, Greenville, NC, United StatesDepartment of Chemistry, East Carolina University, Greenville, NC, United StatesDepartment of Physical Therapy, East Carolina University, Greenville, NC, United StatesDepartment of Physical Therapy, East Carolina University, Greenville, NC, United StatesDepartment of Chemistry, East Carolina University, Greenville, NC, United StatesDepartment of Chemistry, East Carolina University, Greenville, NC, United StatesDepartment of Physical Therapy, East Carolina University, Greenville, NC, United StatesDepartment of Chemistry, East Carolina University, Greenville, NC, United StatesCofilin-actin rods are a well-documented stress response in neuronal cells and their persistence is frequently associated with neurodegenerative disease. However, the role of specific actin residues in promoting the formation of cofilin-actin rods and other anomalous cytoskeletal structures is largely unknown. As it is increasingly suspected that specific mutations and post-translation modifications of actin may promote neurodegenerative disease, characterizing the role of these residues in cytoskeletal dysregulation is highly relevant. In this study, we focus on the actin-ATP interface, which has been proposed as a key mediator of cofilin-actin rod formation and the propensity of actin to respond to cellular stress. Using a light and stress-gated reporter of cofilin-actin cluster formation, we determine the impact of mutants associated with Actin-ATP binding on the propensity of actin to form anomalous structures in the presence and absence of applied cellular stress. This study identifies actin mutants that promote anomalous actin inclusions in HeLa cells and characterizes the manifestation of these phenotypes in cortical neurons. Mutations to the ATP phosphate tail-binding region of actin (K18A, D154A, G158L, K213A) were found to be particularly disruptive to actin phenotypes, and in several instances promote disease-associated actin-rich structures such as cofilin-actin rods and Hirano bodies. We find that these mutant phenotypes are largely consistent between cell types and display highly unusual inclusions in cultured cortical neurons, without leading to nuclear fragmentation and apoptotic death of the transfected cells. These mutants strengthen the association of residue-specific changes in actin with large-scale phenotypic and functional changes in the cytoskeleton, further implicating them in neurodegenerative disease progression.https://www.frontiersin.org/articles/10.3389/fncel.2025.1543199/fullneurodegenerationcytoskeletoncofilin-actin rodsHirano bodiesCry2/CIB1cellular stress |
| spellingShingle | Noah Mann Keerthana Surabhi Josephine Sharp Mary Phipps Maelee Becton Jahiem Hill Davis Roberts Erzsebet M. Szatmari Robert M. Hughes Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface Frontiers in Cellular Neuroscience neurodegeneration cytoskeleton cofilin-actin rods Hirano bodies Cry2/CIB1 cellular stress |
| title | Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface |
| title_full | Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface |
| title_fullStr | Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface |
| title_full_unstemmed | Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface |
| title_short | Identification of actin mutants with neurodegenerative disease-like phenotypes via mutagenesis of the actin-ATP interface |
| title_sort | identification of actin mutants with neurodegenerative disease like phenotypes via mutagenesis of the actin atp interface |
| topic | neurodegeneration cytoskeleton cofilin-actin rods Hirano bodies Cry2/CIB1 cellular stress |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2025.1543199/full |
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