Structural instability of lamin A tail domain modulates its assembly and higher order function in Emery-Dreifuss muscular dystrophy

Biochem Biophys Res Commun. 2019 Apr 23;512(1):22-28. doi: 10.1016/j.bbrc.2019.02.138. Epub 2019 Mar 7.

Abstract

The C-terminal Ig-domain of lamin A plays critical roles in cell function via interaction with proteins, DNA, and chromatin. Mutations in this domain are known to cause various diseases including Emery-Dreifuss muscular dystrophy (EDMD) and familial partial lipodystrophy (FPLD). Here we examined the biophysical and biochemical properties of mutant Ig-domains identified in patients with EDMD and FPLD. EDMD-related mutant Ig-domain showed decreased stability to heat and denaturant. This result was also confirmed by experiments using full-length mutant lamin A, although the decrease in melting temperature was much less than that of the mutant Ig-domain alone. The unstable EDMD Ig-domain disrupted the proper assembly of lamin A, resulting in abnormal paracrystal formation and decreased viscosity. In contrast, FPLD-related mutant Ig-domains were thermally stable, although they lost DNA binding function. Alanine substitution experiments revealed a functional domain of DNA binding in the Ig-domain. Thus, the overall biophysical property of Ig-domains is closely associated with clinical phenotype.

Keywords: Ig-domain; Lamin A; Laminopathy; Thermal stability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Biophysical Phenomena
  • DNA / chemistry
  • DNA / metabolism
  • Humans
  • In Vitro Techniques
  • Lamin Type A / chemistry*
  • Lamin Type A / genetics
  • Lamin Type A / metabolism
  • Lipodystrophy, Familial Partial / genetics
  • Lipodystrophy, Familial Partial / metabolism
  • Microscopy, Electron, Transmission
  • Models, Molecular
  • Muscular Dystrophy, Emery-Dreifuss / genetics
  • Muscular Dystrophy, Emery-Dreifuss / metabolism*
  • Mutation
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / metabolism
  • Protein Domains
  • Protein Stability
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Lamin Type A
  • Peptide Fragments
  • Recombinant Proteins
  • DNA