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LMNA variants cause cytoplasmic distribution of nuclear pore proteins in Drosophila and human muscle

By George Dialynas, Kaitlin M. Flannery, Luka N. Zirbel, Peter L. Nagy, Katherine D. Mathews, Steven A. Moore and Lori L. Wallrath

Abstract

Mutations in the human LMNA gene, encoding A-type lamins, give rise to laminopathies, which include several types of muscular dystrophy. Here, heterozygous sequence variants in LMNA, which result in single amino-acid substitutions, were identified in patients exhibiting muscle weakness. To assess whether the substitutions altered lamin function, we performed in vivo analyses using a Drosophila model. Stocks were generated that expressed mutant forms of the Drosophila A-type lamin modeled after each variant. Larvae were used for motility assays and histochemical staining of the body-wall muscle. In parallel, immunohistochemical analyses were performed on human muscle biopsy samples from the patients. In control flies, muscle-specific expression of the wild-type A-type lamin had no apparent affect. In contrast, expression of the mutant A-type lamins caused dominant larval muscle defects and semi-lethality at the pupal stage. Histochemical staining of larval body wall muscle revealed that the mutant A-type lamin, B-type lamins, the Sad1p, UNC-84 domain protein Klaroid and nuclear pore complex proteins were mislocalized to the cytoplasm. In addition, cytoplasmic actin filaments were disorganized, suggesting links between the nuclear lamina and the cytoskeleton were disrupted. Muscle biopsies from the patients showed dystrophic histopathology and architectural abnormalities similar to the Drosophila larvae, including cytoplasmic distribution of nuclear envelope proteins. These data provide evidence that the Drosophila model can be used to assess the function of novel LMNA mutations and support the idea that loss of cellular compartmentalization of nuclear proteins contributes to muscle disease pathogenesis

Topics: Articles
Publisher: Oxford University Press
OAI identifier: oai:pubmedcentral.nih.gov:3298278
Provided by: PubMed Central

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Citations

  1. (1986). A cell free system to study reassembly of the nuclear envelope at the end of mitosis.
  2. (2009). A comparative study of Drosophila and human A-type lamins.
  3. (2002). A novel interaction between lamin A and SREBP1: implications for partial lipodystrophy and other laminopathies.
  4. (2006). A-type lamin complexes and regenerative potential: a step towards understanding laminopathic diseases?
  5. (2004). A-type lamins regulate retinoblastoma protein function by promoting subnuclear localization and preventing proteasomal degradation.
  6. (2008). Adult stem cell maintenance and tissue regeneration in the ageing context: the role for A-type lamins as intrinsic modulators of ageing in adult stem cells and their niches.
  7. (1996). An alternative splicing product of the lamin A/C gene lacks exon 10.
  8. (2003). Architectural abnormalities in muscle nuclei: ultrastructural differences between X-linked and autosomal dominant forms of EDMD.
  9. (1980). Arrangement of chromatin in the nucleus.
  10. (1997). Assembly of A- and B-type lamins studied in vivo with the baculovirus system.
  11. (1993). cDNA cloning of a germ cell specific lamin B3 from mouse spermatocytes and analysis of its function by ectopic expression in somatic cells.
  12. (1986). cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins.
  13. (2006). Coupling of the nucleus and cytoplasm: role of the LINC complex.
  14. (2006). Dependence of diffusional mobility of integral inner nuclear membrane proteins on A-type lamins.
  15. (2011). Distinct association of the nuclear pore protein Nup153 with A- and B-type lamins.
  16. (1989). Distribution patterns of HP1, a heterochromatin-associated nonhistone chromosomal protein of Drosophila.
  17. (2007). Drosophila Klaroid encodes a SUN domain protein required for Klarsicht localization to the nuclear envelope and nuclear migration in the eye.
  18. (2007). dTrf2 is required for transcriptional and developmental responses to ecdysone during Drosophila metamorphosis.
  19. (2000). Dynamic associations of heterochromatin protein 1 with the nuclear envelope.
  20. (2000). Essential roles for Caenorhabditis elegans lamin gene in nuclear organization, cell cycle progression, and spatial organization of nuclear pore complexes.
  21. (1995). Expression of Drosophila lamin C is developmentally regulated: analogies with vertebrate A-type lamins.
  22. (1988). Functional organization of the nuclear envelope.
  23. (2002). GAL4 system in Drosophila: a fly geneticist’s Swiss army knife.
  24. (1996). Genetic modification of heterochromatic association and nuclear organization in Drosophila.
  25. (2010). Hereditary muscular dystrophies and the heart.
  26. (2006). Human laminopathies: nuclei gone genetically awry.
  27. (1994). Identification and cloning of an mRNA coding for a germ cell-specific A-type lamin in mice.
  28. (2006). Identification of a novel, highly variable aminoterminal amino acid sequence element in the nuclear intermediate filament protein lamin B(2) from higher vertebrates.
  29. (2001). Identification of essential genes in cultured mammalian cells using small interfering RNAs.
  30. (1990). In vitro posttranslational modification of lamin B cloned from a human T-cell line.
  31. (1990). Interphase nuclear envelope lamins form a discontinuous network that interacts with only a fraction of the chromatin in the nuclear periphery.
  32. (1999). Intracellular trafficking of emerin, the Emery– Dreifuss muscular dystrophy protein.
  33. (2007). Invertebrate lamins.
  34. (2005). KASH ’n Karry: the KASH domain family of cargo-specific cytoskeletal adaptor proteins.
  35. (2006). KASH-domain proteins in nuclear migration, anchorage and other processes.
  36. (1995). Lamin proteins form an internal nucleoskeleton as well as a peripheral lamina in human cells.
  37. (2010). Lamin-binding proteins.
  38. (2007). Lamins and lamin-associated proteins in aging and disease.
  39. (1994). Lamins and lamin-associated proteins.
  40. (1999). Loss of A-type lamin expression compromises nuclear envelope integrity leading to muscular dystrophy.
  41. (1993). Macromolecular domains within the cell nucleus.
  42. (2006). Methylation-independent binding to histone H3 and cell cycle-dependent incorporation of HP1beta into heterochromatin.
  43. (1983). Mitotic architecture of the cell: the filament networks of the nucleus and cytoplasm.
  44. (2005). Molecular genetic analysis of the nested Drosophila melanogaster lamin C gene.
  45. (1985). Monoclonal antibodies prepared against the major Drosophila nuclear Matrix-pore complex-lamina glycoprotein bind specifically to the nuclear envelope in situ.
  46. (1990). Morphological plasticity of motor axons in Drosophila mutants with altered excitability.
  47. (2006). Mutation Glu82Lys in lamin A/C gene is associated with cardiomyopathy and conduction defect.
  48. (1999). Mutations in the gene encoding lamin A/C cause autosomal dominant Emery–Dreifuss muscular dystrophy.
  49. (2001). Novel and recurrent mutations in lamin A/C in patients with Emery–Dreifuss muscular dystrophy.
  50. (2011). Novel LMNA mutations in patients with Emery–Dreifuss muscular dystrophy and functional characterization of four LMNA mutations.
  51. (1998). Nuclear lamins: their structure, assembly, and interactions.
  52. (1997). Nuclear membrane dynamics and reassembly in living cells: targeting of an inner nuclear membrane protein in interphase and mitosis.
  53. (2003). Nuclear membrane proteins with potential disease links found by subtractive proteomics.
  54. (2000). Nuclear organization of DNA replication in primary mammalian cells.
  55. (1987). Nuclear pore complex contains a family of glycoproteins that includes p62: glycosylation through a previously unidentified cellular pathway.
  56. (2003). Nuclear positioning: the means is at the ends.
  57. (2007). Nucleoplasmic LAP2alpha-lamin A complexes are required to maintain a proliferative state in human fibroblasts.
  58. (2004). Proteins that bind A-type lamins: integrating isolated clues.
  59. (1994). Raising large quantities of Drosophila for biochemical experiments.
  60. (1999). Regulation of DLG localization at synapses by CaMKII-dependent phosphorylation.
  61. (2011). Repetitive disruptions of the nuclear envelope invoke temporary loss of cellular compartmentalization in laminopathies.
  62. (2007). Role of nuclear lamina–cytoskeleton interactions in the maintenance of cellular strength.
  63. (2008). Sensory mechanisms controlling the timing of larval developmental and behavioral transitions require
  64. (2002). Structure of the globular tail of nuclear lamin.
  65. (2006). SUN1 interacts with nuclear lamin A and cytoplasmic nesprins to provide a physical connection between the nuclear lamina and the cytoskeleton.
  66. (2004). Sun2 is a novel mammalian inner nuclear membrane protein.
  67. (1993). Targeted gene expression as a means of altering cell fates and generating dominant phenotypes.
  68. (1980). The nuclear envelope lamina is reversibly depolymerized during mitosis.
  69. (2004). The nuclear envelope lamina network has elasticity and a compressibility limit suggestive of a molecular shock absorber.
  70. (2007). The nuclear envelope, a key structure in cellular integrity and gene expression.
  71. (2002). The nuclear envelope, lamins and nuclear assembly.
  72. (2011). The nucleoporin Nup88 is interacting with nuclear lamin A.
  73. (2003). The nucleoskeleton: lamins and actin are major players in essential nuclear functions.
  74. (2001). The plant nuclear envelope.
  75. (2010). The role of Drosophila Lamin C in muscle function and gene expression.
  76. (1992). The urate oxidase gene of Drosophila pseudoobscura and Drosophila melanogaster: evolutionary changes of sequence and regulation.
  77. (1998). Ultrastructural abnormality of sarcolemmal nuclei in Emery–Dreifuss muscular dystrophy (EDMD).

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