β-III spectrin mutation L253P associated with spinocerebellar ataxia type 5 interferes with binding to Arp1 and protein trafficking from the Golgi

Yvonne L. Clarkson, Trudi Gillespie, Emma M. Perkins, Alastair R. Lyndon, Mandy Jackson

    Research output: Contribution to journalArticle

    Abstract

    Spinocerebellar ataxia type 5 (SCA5) is an autosomal dominant neurodegenerative disorder caused by mutations in ß-III spectrin. A mouse lacking full-length ß-III spectrin has a phenotype closely mirroring symptoms of SCA5 patients. Here we report the analysis of heterozygous animals, which show no signs of ataxia or cerebellar degeneration up to 2 years of age. This argues against haploinsufficiency as a disease mechanism and points towards human mutations having a dominant-negative effect on wild-type (WT) ß-III spectrin function. Cell culture studies using ß-III spectrin with a mutation associated with SCA5 (L253P) reveal that mutant protein, instead of being found at the cell membrane, appears trapped in the cytoplasm associated with the Golgi apparatus. Furthermore, L253P ß-III spectrin prevents correct localization of WT ß-III spectrin and prevents EAAT4, a protein known to interact with ß-III spectrin, from reaching the plasma membrane. Interaction of ß-III spectrin with Arp1, a subunit of the dynactin-dynein complex, is also lost with the L253P substitution. Despite intracellular accumulation of proteins, this cellular stress does not induce the unfolded protein response, implying the importance of membrane protein loss in disease pathogenesis. Incubation at lower temperature (25°C) rescues L253P ß-III spectrin interaction with Arp1 and normal protein trafficking to the membrane. These data provide evidence for a dominant-negative effect of an SCA5 mutation and show for the first time that trafficking of both ß-III spectrin and EAAT4 from the Golgi is disrupted through failure of the L253P mutation to interact with Arp1. © The Author 2010. Published by Oxford University Press. All rights reserved.

    Original languageEnglish
    Article numberddq279
    Pages (from-to)3634-3641
    Number of pages8
    JournalHuman Molecular Genetics
    Volume19
    Issue number18
    DOIs
    Publication statusPublished - 5 Jul 2010

    Fingerprint

    Spinocerebellar Ataxias
    Spectrin
    Protein Transport
    Mutation
    Cell Membrane
    Unfolded Protein Response
    Haploinsufficiency
    Dyneins
    Cerebellar Ataxia
    Golgi Apparatus
    Mutant Proteins
    Heat-Shock Proteins
    Neurodegenerative Diseases
    Membrane Proteins
    Cytoplasm
    Cell Culture Techniques

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    @article{b355b2467ebd4808b6e2f64bc3473744,
    title = "β-III spectrin mutation L253P associated with spinocerebellar ataxia type 5 interferes with binding to Arp1 and protein trafficking from the Golgi",
    abstract = "Spinocerebellar ataxia type 5 (SCA5) is an autosomal dominant neurodegenerative disorder caused by mutations in {\ss}-III spectrin. A mouse lacking full-length {\ss}-III spectrin has a phenotype closely mirroring symptoms of SCA5 patients. Here we report the analysis of heterozygous animals, which show no signs of ataxia or cerebellar degeneration up to 2 years of age. This argues against haploinsufficiency as a disease mechanism and points towards human mutations having a dominant-negative effect on wild-type (WT) {\ss}-III spectrin function. Cell culture studies using {\ss}-III spectrin with a mutation associated with SCA5 (L253P) reveal that mutant protein, instead of being found at the cell membrane, appears trapped in the cytoplasm associated with the Golgi apparatus. Furthermore, L253P {\ss}-III spectrin prevents correct localization of WT {\ss}-III spectrin and prevents EAAT4, a protein known to interact with {\ss}-III spectrin, from reaching the plasma membrane. Interaction of {\ss}-III spectrin with Arp1, a subunit of the dynactin-dynein complex, is also lost with the L253P substitution. Despite intracellular accumulation of proteins, this cellular stress does not induce the unfolded protein response, implying the importance of membrane protein loss in disease pathogenesis. Incubation at lower temperature (25°C) rescues L253P {\ss}-III spectrin interaction with Arp1 and normal protein trafficking to the membrane. These data provide evidence for a dominant-negative effect of an SCA5 mutation and show for the first time that trafficking of both {\ss}-III spectrin and EAAT4 from the Golgi is disrupted through failure of the L253P mutation to interact with Arp1. {\circledC} The Author 2010. Published by Oxford University Press. All rights reserved.",
    author = "Clarkson, {Yvonne L.} and Trudi Gillespie and Perkins, {Emma M.} and Lyndon, {Alastair R.} and Mandy Jackson",
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    β-III spectrin mutation L253P associated with spinocerebellar ataxia type 5 interferes with binding to Arp1 and protein trafficking from the Golgi. / Clarkson, Yvonne L.; Gillespie, Trudi; Perkins, Emma M.; Lyndon, Alastair R.; Jackson, Mandy.

    In: Human Molecular Genetics, Vol. 19, No. 18, ddq279, 05.07.2010, p. 3634-3641.

    Research output: Contribution to journalArticle

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    AU - Clarkson, Yvonne L.

    AU - Gillespie, Trudi

    AU - Perkins, Emma M.

    AU - Lyndon, Alastair R.

    AU - Jackson, Mandy

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    N2 - Spinocerebellar ataxia type 5 (SCA5) is an autosomal dominant neurodegenerative disorder caused by mutations in ß-III spectrin. A mouse lacking full-length ß-III spectrin has a phenotype closely mirroring symptoms of SCA5 patients. Here we report the analysis of heterozygous animals, which show no signs of ataxia or cerebellar degeneration up to 2 years of age. This argues against haploinsufficiency as a disease mechanism and points towards human mutations having a dominant-negative effect on wild-type (WT) ß-III spectrin function. Cell culture studies using ß-III spectrin with a mutation associated with SCA5 (L253P) reveal that mutant protein, instead of being found at the cell membrane, appears trapped in the cytoplasm associated with the Golgi apparatus. Furthermore, L253P ß-III spectrin prevents correct localization of WT ß-III spectrin and prevents EAAT4, a protein known to interact with ß-III spectrin, from reaching the plasma membrane. Interaction of ß-III spectrin with Arp1, a subunit of the dynactin-dynein complex, is also lost with the L253P substitution. Despite intracellular accumulation of proteins, this cellular stress does not induce the unfolded protein response, implying the importance of membrane protein loss in disease pathogenesis. Incubation at lower temperature (25°C) rescues L253P ß-III spectrin interaction with Arp1 and normal protein trafficking to the membrane. These data provide evidence for a dominant-negative effect of an SCA5 mutation and show for the first time that trafficking of both ß-III spectrin and EAAT4 from the Golgi is disrupted through failure of the L253P mutation to interact with Arp1. © The Author 2010. Published by Oxford University Press. All rights reserved.

    AB - Spinocerebellar ataxia type 5 (SCA5) is an autosomal dominant neurodegenerative disorder caused by mutations in ß-III spectrin. A mouse lacking full-length ß-III spectrin has a phenotype closely mirroring symptoms of SCA5 patients. Here we report the analysis of heterozygous animals, which show no signs of ataxia or cerebellar degeneration up to 2 years of age. This argues against haploinsufficiency as a disease mechanism and points towards human mutations having a dominant-negative effect on wild-type (WT) ß-III spectrin function. Cell culture studies using ß-III spectrin with a mutation associated with SCA5 (L253P) reveal that mutant protein, instead of being found at the cell membrane, appears trapped in the cytoplasm associated with the Golgi apparatus. Furthermore, L253P ß-III spectrin prevents correct localization of WT ß-III spectrin and prevents EAAT4, a protein known to interact with ß-III spectrin, from reaching the plasma membrane. Interaction of ß-III spectrin with Arp1, a subunit of the dynactin-dynein complex, is also lost with the L253P substitution. Despite intracellular accumulation of proteins, this cellular stress does not induce the unfolded protein response, implying the importance of membrane protein loss in disease pathogenesis. Incubation at lower temperature (25°C) rescues L253P ß-III spectrin interaction with Arp1 and normal protein trafficking to the membrane. These data provide evidence for a dominant-negative effect of an SCA5 mutation and show for the first time that trafficking of both ß-III spectrin and EAAT4 from the Golgi is disrupted through failure of the L253P mutation to interact with Arp1. © The Author 2010. Published by Oxford University Press. All rights reserved.

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