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

Hum Mol Genet. 2012 Apr 1;21(7):1544-56. doi: 10.1093/hmg/ddr592. Epub 2011 Dec 20.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Child
  • Child, Preschool
  • Cytoplasm / chemistry
  • Drosophila / genetics
  • Drosophila / growth & development
  • Drosophila Proteins / analysis
  • Drosophila Proteins / chemistry
  • Drosophila Proteins / genetics*
  • Genetic Variation
  • Humans
  • Lamin Type A / analysis
  • Lamin Type A / chemistry
  • Lamin Type A / genetics*
  • Lamin Type B / analysis
  • Models, Molecular
  • Molecular Sequence Data
  • Motor Activity
  • Muscle Weakness / genetics
  • Muscle, Skeletal / chemistry*
  • Muscle, Skeletal / pathology
  • Muscular Atrophy / genetics
  • Muscular Atrophy / pathology
  • Muscular Dystrophies / genetics*
  • Muscular Dystrophies / pathology
  • Nuclear Pore Complex Proteins / analysis*
  • Protein Structure, Tertiary / genetics

Substances

  • Drosophila Proteins
  • LMNA protein, human
  • Lamin Type A
  • Lamin Type B
  • Nuclear Pore Complex Proteins