Conserved role for Ataxin-2 in mediating endoplasmic reticulum dynamics

Urko del Castillo; Megan M. Gnazzo; Christopher G. Sorensen Turpin; Ken C.Q. Nguyen; Emily Semaya; Yuwan Lam; Matthew A. de Cruz; Joshua N. Bembenek; David H. Hall; Blake Riggs; Vladimir I. Gelfand; Ahna R. Skop

doi:10.1111/tra.12647

Conserved role for Ataxin-2 in mediating endoplasmic reticulum dynamics

Urko del Castillo, Megan M. Gnazzo, Christopher G. Sorensen Turpin, Ken C.Q. Nguyen, Emily Semaya, Yuwan Lam, Matthew A. de Cruz, Joshua N. Bembenek, David H. Hall, Blake Riggs, Vladimir I. Gelfand, Ahna R. Skop^*

^*Corresponding author for this work

Cell & Developmental Biology

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

Ataxin-2, a conserved RNA-binding protein, is implicated in the late-onset neurodegenerative disease Spinocerebellar ataxia type-2 (SCA2). SCA2 is characterized by shrunken dendritic arbors and torpedo-like axons within the Purkinje neurons of the cerebellum. Torpedo-like axons have been described to contain displaced endoplasmic reticulum (ER) in the periphery of the cell; however, the role of Ataxin-2 in mediating ER function in SCA2 is unclear. We utilized the Caenorhabditis elegans and Drosophila homologs of Ataxin-2 (ATX-2 and DAtx2, respectively) to determine the role of Ataxin-2 in ER function and dynamics in embryos and neurons. Loss of ATX-2 and DAtx2 resulted in collapse of the ER in dividing embryonic cells and germline, and ultrastructure analysis revealed unique spherical stacks of ER in mature oocytes and fragmented and truncated ER tubules in the embryo. ATX-2 and DAtx2 reside in puncta adjacent to the ER in both C. elegans and Drosophila embryos. Lastly, depletion of DAtx2 in cultured Drosophila neurons recapitulated the shrunken dendritic arbor phenotype of SCA2. ER morphology and dynamics were severely disrupted in these neurons. Taken together, we provide evidence that Ataxin-2 plays an evolutionary conserved role in ER dynamics and morphology in C. elegans and Drosophila embryos during development and in fly neurons, suggesting a possible SCA2 disease mechanism.

Original language	English (US)
Pages (from-to)	436-447
Number of pages	12
Journal	Traffic
Volume	20
Issue number	6
DOIs	https://doi.org/10.1111/tra.12647
State	Published - Jun 2019

Funding

We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenbüechler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943. National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 information National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenb?echler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943.

Keywords

ATX-2
Ataxin-2
DAtx-2
RBPs
RNA
cytokinesis
endoplasmic reticulum
mitosis
neurons

ASJC Scopus subject areas

Genetics
Molecular Biology
Structural Biology
Biochemistry
Cell Biology

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10.1111/tra.12647

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@article{377fa54d3dbf43d6aaab50fb9bb059cf,

title = "Conserved role for Ataxin-2 in mediating endoplasmic reticulum dynamics",

abstract = "Ataxin-2, a conserved RNA-binding protein, is implicated in the late-onset neurodegenerative disease Spinocerebellar ataxia type-2 (SCA2). SCA2 is characterized by shrunken dendritic arbors and torpedo-like axons within the Purkinje neurons of the cerebellum. Torpedo-like axons have been described to contain displaced endoplasmic reticulum (ER) in the periphery of the cell; however, the role of Ataxin-2 in mediating ER function in SCA2 is unclear. We utilized the Caenorhabditis elegans and Drosophila homologs of Ataxin-2 (ATX-2 and DAtx2, respectively) to determine the role of Ataxin-2 in ER function and dynamics in embryos and neurons. Loss of ATX-2 and DAtx2 resulted in collapse of the ER in dividing embryonic cells and germline, and ultrastructure analysis revealed unique spherical stacks of ER in mature oocytes and fragmented and truncated ER tubules in the embryo. ATX-2 and DAtx2 reside in puncta adjacent to the ER in both C. elegans and Drosophila embryos. Lastly, depletion of DAtx2 in cultured Drosophila neurons recapitulated the shrunken dendritic arbor phenotype of SCA2. ER morphology and dynamics were severely disrupted in these neurons. Taken together, we provide evidence that Ataxin-2 plays an evolutionary conserved role in ER dynamics and morphology in C. elegans and Drosophila embryos during development and in fly neurons, suggesting a possible SCA2 disease mechanism.",

keywords = "ATX-2, Ataxin-2, DAtx-2, RBPs, RNA, cytokinesis, endoplasmic reticulum, mitosis, neurons",

author = "{del Castillo}, Urko and Gnazzo, {Megan M.} and {Sorensen Turpin}, {Christopher G.} and Nguyen, {Ken C.Q.} and Emily Semaya and Yuwan Lam and {de Cruz}, {Matthew A.} and Bembenek, {Joshua N.} and Hall, {David H.} and Blake Riggs and Gelfand, {Vladimir I.} and Skop, {Ahna R.}",

note = "Funding Information: We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenb{\"u}echler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943. Funding Information: National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 Funding Information: information National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenb?echler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943. Publisher Copyright: {\textcopyright} 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd",

year = "2019",

month = jun,

doi = "10.1111/tra.12647",

language = "English (US)",

volume = "20",

pages = "436--447",

journal = "Traffic",

issn = "1398-9219",

publisher = "Blackwell Munksgaard",

number = "6",

}

TY - JOUR

T1 - Conserved role for Ataxin-2 in mediating endoplasmic reticulum dynamics

AU - del Castillo, Urko

AU - Gnazzo, Megan M.

AU - Sorensen Turpin, Christopher G.

AU - Nguyen, Ken C.Q.

AU - Semaya, Emily

AU - Lam, Yuwan

AU - de Cruz, Matthew A.

AU - Bembenek, Joshua N.

AU - Hall, David H.

AU - Riggs, Blake

AU - Gelfand, Vladimir I.

AU - Skop, Ahna R.

N1 - Funding Information: We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenbüechler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943. Funding Information: National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 Funding Information: information National Science Foundation, Grant/Award Numbers: MCB 1158003, CAREER MCB 1553695, MRI 0821204; National Institutes of Health, Grant/Award Numbers: R01 GM114471, R01 GM52111, OD 010943; National Cancer Institute, Grant/Award Number: CCSG P30 CA060553; National Institutes of Health Office of Research Infrastructure Programs, Grant/Award Number: P40 OD010440; NSF MRI 0821204 We thank Randall Dahn for advice and for critically reading the manuscript, Jill Willdonger for initially performing DAtx2 knockdown experiments in sensory neurons, and David Kirchenb?echler (Northwestern University Center for Advanced Microscopy), for help with ER FRAP quantification. Drosophila Stocks obtained from the Bloomington Drosophila Stock Center (NIH P40OD018537) were used in this study. Some Caenorhabditis elegans strains above were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). A.R.S. was supported by the National Science Foundation under award MCB 1158003. C. elegans strains were obtained from the C. elegans Genetics Stock Center which is supported by the National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). J.N.B. was supported by NIH R01 GM114471. V.I.G was supported by the National Institute of General Medical Sciences of the National Institutes of Health under award R01 GM52111. The imaging work that was performed at the Northwestern University Center for Advanced Microscopy generously supported by NCI CCSG P30 CA060553 awarded to the Robert H Lurie Comprehensive Cancer Center. B.R. was supported by the National Science Foundation CAREER Award 1553695. Imaging work involving the early Drosophila embryo was performed at the San Francisco State University, Cell and Molecular Imaging Center (CMIC) with the generous assistance of Annette Chan. D.H.H. was supported by NIH OD 010943. Publisher Copyright: © 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd

PY - 2019/6

Y1 - 2019/6

N2 - Ataxin-2, a conserved RNA-binding protein, is implicated in the late-onset neurodegenerative disease Spinocerebellar ataxia type-2 (SCA2). SCA2 is characterized by shrunken dendritic arbors and torpedo-like axons within the Purkinje neurons of the cerebellum. Torpedo-like axons have been described to contain displaced endoplasmic reticulum (ER) in the periphery of the cell; however, the role of Ataxin-2 in mediating ER function in SCA2 is unclear. We utilized the Caenorhabditis elegans and Drosophila homologs of Ataxin-2 (ATX-2 and DAtx2, respectively) to determine the role of Ataxin-2 in ER function and dynamics in embryos and neurons. Loss of ATX-2 and DAtx2 resulted in collapse of the ER in dividing embryonic cells and germline, and ultrastructure analysis revealed unique spherical stacks of ER in mature oocytes and fragmented and truncated ER tubules in the embryo. ATX-2 and DAtx2 reside in puncta adjacent to the ER in both C. elegans and Drosophila embryos. Lastly, depletion of DAtx2 in cultured Drosophila neurons recapitulated the shrunken dendritic arbor phenotype of SCA2. ER morphology and dynamics were severely disrupted in these neurons. Taken together, we provide evidence that Ataxin-2 plays an evolutionary conserved role in ER dynamics and morphology in C. elegans and Drosophila embryos during development and in fly neurons, suggesting a possible SCA2 disease mechanism.

AB - Ataxin-2, a conserved RNA-binding protein, is implicated in the late-onset neurodegenerative disease Spinocerebellar ataxia type-2 (SCA2). SCA2 is characterized by shrunken dendritic arbors and torpedo-like axons within the Purkinje neurons of the cerebellum. Torpedo-like axons have been described to contain displaced endoplasmic reticulum (ER) in the periphery of the cell; however, the role of Ataxin-2 in mediating ER function in SCA2 is unclear. We utilized the Caenorhabditis elegans and Drosophila homologs of Ataxin-2 (ATX-2 and DAtx2, respectively) to determine the role of Ataxin-2 in ER function and dynamics in embryos and neurons. Loss of ATX-2 and DAtx2 resulted in collapse of the ER in dividing embryonic cells and germline, and ultrastructure analysis revealed unique spherical stacks of ER in mature oocytes and fragmented and truncated ER tubules in the embryo. ATX-2 and DAtx2 reside in puncta adjacent to the ER in both C. elegans and Drosophila embryos. Lastly, depletion of DAtx2 in cultured Drosophila neurons recapitulated the shrunken dendritic arbor phenotype of SCA2. ER morphology and dynamics were severely disrupted in these neurons. Taken together, we provide evidence that Ataxin-2 plays an evolutionary conserved role in ER dynamics and morphology in C. elegans and Drosophila embryos during development and in fly neurons, suggesting a possible SCA2 disease mechanism.

KW - ATX-2

KW - Ataxin-2

KW - DAtx-2

KW - RBPs

KW - RNA

KW - cytokinesis

KW - endoplasmic reticulum

KW - mitosis

KW - neurons

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DO - 10.1111/tra.12647

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AN - SCOPUS:85065669792

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VL - 20

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JO - Traffic

JF - Traffic

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ER -

Conserved role for Ataxin-2 in mediating endoplasmic reticulum dynamics

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