The ZZ domain of dystrophin in DMD: Making sense of missense mutations

Adeline Vulin, Nicolas Wein, Dana M. Strandjord, Eric K. Johnson, Andrew R. Findlay, Baijayanta Maiti, Michael T. Howard, Yuuki J. Kaminoh, Laura E. Taylor, Tabatha R. Simmons, Will C. Ray, Federica Montanaro, James M Ervasti, Kevin M. Flanigan

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Duchenne muscular dystrophy (DMD) is associated with the loss of dystrophin, which plays an important role in myofiber integrity via interactions with β-dystroglycan and other members of the transmembrane dystrophin-associated protein complex. The ZZ domain, a cysteine-rich zinc-finger domain near the dystrophin C-terminus, is implicated in forming a stable interaction between dystrophin and β-dystroglycan, but the mechanism of pathogenesis of ZZ missense mutations has remained unclear because not all such mutations have been shown to alter β-dystroglycan binding in previous experimental systems. We engineered three ZZ mutations (p.Cys3313Phe, p.Asp3335His, and p.Cys3340Tyr) into a short construct similar to the Dp71 dystrophin isoform for in vitro and in vivo studies and delineated their effect on protein expression, folding properties, and binding partners. Our results demonstrate two distinct pathogenic mechanisms for ZZ missense mutations. The cysteine mutations result in diminished or absent subsarcolemmal expression because of protein instability, likely due to misfolding. In contrast, the aspartic acid mutation disrupts binding with β-dystroglycan despite an almost normal expression at the membrane, confirming a role for the ZZ domain in β-dystroglycan binding but surprisingly demonstrating that such binding is not required for subsarcolemmal localization of dystrophin, even in the absence of actin binding domains.

Original languageEnglish (US)
Pages (from-to)257-264
Number of pages8
JournalHuman mutation
Volume35
Issue number2
DOIs
StatePublished - Feb 1 2014

Fingerprint

Dystroglycans
Dystrophin
Duchenne Muscular Dystrophy
Missense Mutation
Mutation
Cysteine
Dystrophin-Associated Protein Complex
Protein Folding
Zinc Fingers
Aspartic Acid
Actins
Protein Isoforms
Membranes
Proteins

Keywords

  • Duchenne muscular dystrophy
  • Dystrophin
  • Missense mutation
  • ZZ domain

Cite this

Vulin, A., Wein, N., Strandjord, D. M., Johnson, E. K., Findlay, A. R., Maiti, B., ... Flanigan, K. M. (2014). The ZZ domain of dystrophin in DMD: Making sense of missense mutations. Human mutation, 35(2), 257-264. https://doi.org/10.1002/humu.22479

The ZZ domain of dystrophin in DMD : Making sense of missense mutations. / Vulin, Adeline; Wein, Nicolas; Strandjord, Dana M.; Johnson, Eric K.; Findlay, Andrew R.; Maiti, Baijayanta; Howard, Michael T.; Kaminoh, Yuuki J.; Taylor, Laura E.; Simmons, Tabatha R.; Ray, Will C.; Montanaro, Federica; Ervasti, James M; Flanigan, Kevin M.

In: Human mutation, Vol. 35, No. 2, 01.02.2014, p. 257-264.

Research output: Contribution to journalArticle

Vulin, A, Wein, N, Strandjord, DM, Johnson, EK, Findlay, AR, Maiti, B, Howard, MT, Kaminoh, YJ, Taylor, LE, Simmons, TR, Ray, WC, Montanaro, F, Ervasti, JM & Flanigan, KM 2014, 'The ZZ domain of dystrophin in DMD: Making sense of missense mutations', Human mutation, vol. 35, no. 2, pp. 257-264. https://doi.org/10.1002/humu.22479
Vulin A, Wein N, Strandjord DM, Johnson EK, Findlay AR, Maiti B et al. The ZZ domain of dystrophin in DMD: Making sense of missense mutations. Human mutation. 2014 Feb 1;35(2):257-264. https://doi.org/10.1002/humu.22479
Vulin, Adeline ; Wein, Nicolas ; Strandjord, Dana M. ; Johnson, Eric K. ; Findlay, Andrew R. ; Maiti, Baijayanta ; Howard, Michael T. ; Kaminoh, Yuuki J. ; Taylor, Laura E. ; Simmons, Tabatha R. ; Ray, Will C. ; Montanaro, Federica ; Ervasti, James M ; Flanigan, Kevin M. / The ZZ domain of dystrophin in DMD : Making sense of missense mutations. In: Human mutation. 2014 ; Vol. 35, No. 2. pp. 257-264.
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AU - Maiti, Baijayanta

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AU - Simmons, Tabatha R.

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AB - Duchenne muscular dystrophy (DMD) is associated with the loss of dystrophin, which plays an important role in myofiber integrity via interactions with β-dystroglycan and other members of the transmembrane dystrophin-associated protein complex. The ZZ domain, a cysteine-rich zinc-finger domain near the dystrophin C-terminus, is implicated in forming a stable interaction between dystrophin and β-dystroglycan, but the mechanism of pathogenesis of ZZ missense mutations has remained unclear because not all such mutations have been shown to alter β-dystroglycan binding in previous experimental systems. We engineered three ZZ mutations (p.Cys3313Phe, p.Asp3335His, and p.Cys3340Tyr) into a short construct similar to the Dp71 dystrophin isoform for in vitro and in vivo studies and delineated their effect on protein expression, folding properties, and binding partners. Our results demonstrate two distinct pathogenic mechanisms for ZZ missense mutations. The cysteine mutations result in diminished or absent subsarcolemmal expression because of protein instability, likely due to misfolding. In contrast, the aspartic acid mutation disrupts binding with β-dystroglycan despite an almost normal expression at the membrane, confirming a role for the ZZ domain in β-dystroglycan binding but surprisingly demonstrating that such binding is not required for subsarcolemmal localization of dystrophin, even in the absence of actin binding domains.

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