Caveolin-3 T78M and T78K missense mutations lead to different phenotypes in vivo and in vitro

Monica Traverso, Elisabetta Gazzerro, Stefania Assereto, Federica Sotgia, Roberta Biancheri, Silvia Stringara, Laura Giberti, Marina Pedemonte, Xiabo Wang, Sara Scapolan, Elisabetta Pasquini, Maria A. Donati, Federico Zara, Michael P. Lisanti, Claudio Bruno, Carlo Minetti

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Caveolins are the principal protein components of caveolae, invaginations of the plasma membrane involved in cell signaling and trafficking. Caveolin-3 (Cav-3) is the muscle-specific isoform of the caveolin family and mutations in the CAV3 gene lead to a large group of neuromuscular disorders. In unrelated patients, we identified two distinct CAV3 mutations involving the same codon 78. Patient 1, affected by dilated cardiomyopathy and limb girdle muscular dystrophy (LGMD)-1C, shows an autosomal recessive mutation converting threonine to methionine (T78M). Patient 2, affected by isolated familiar hyperCKemia, shows an autosomal dominant mutation converting threonine to lysine (T78K). Cav-3 wild type (WT) and Cav-3 mutations were transiently transfected into Cos-7 cells. Cav-3 WT and Cav-3 T78M mutant localized at the plasma membrane, whereas Cav-3 T78K was retained in a perinuclear compartment. Cav-3 T78K expression was decreased by 87% when compared with Cav-3 WT, whereas Cav-3 T78M protein levels were unchanged. To evaluate whether Cav-3 T78K and Cav-3 T78M mutants behaved with a dominant negative pattern, Cos-7 cells were cotransfected with green fluorescent protein (GFP)-Cav-3 WT in combination with either mutant or WT Cav-3. When cotransfected with Cav-3 WT or Cav-3 T78M, GFP-Cav-3 WT was localized at the plasma membrane, as expected. However, when cotransfected with Cav-3 T78K, GFP-Cav-3 WT was retained in a perinuclear compartment, and its protein levels were reduced by 60%, suggesting a dominant negative action. Accordingly, Cav-3 protein levels in muscles from a biopsy of patient 2 (T78K mutation) were reduced by 80%. In conclusion, CAV3 T78M and T78K mutations lead to distinct disorders showing different clinical features and inheritance, and displaying distinct phenotypes in vitro.

Original languageEnglish
Pages (from-to)275-283
Number of pages9
JournalLaboratory Investigation
Volume88
Issue number3
DOIs
Publication statusPublished - Mar 2008

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Caveolin 3
Missense Mutation
Phenotype
Mutation
Caveolins
Green Fluorescent Proteins
In Vitro Techniques
Cell Membrane
Threonine

Keywords

  • Caveolin-3
  • Dilated cardiomyopathy
  • Hyperckemia
  • Hypertrophic cardiomyopathy
  • Limb girdle muscular dystrophy

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Caveolin-3 T78M and T78K missense mutations lead to different phenotypes in vivo and in vitro. / Traverso, Monica; Gazzerro, Elisabetta; Assereto, Stefania; Sotgia, Federica; Biancheri, Roberta; Stringara, Silvia; Giberti, Laura; Pedemonte, Marina; Wang, Xiabo; Scapolan, Sara; Pasquini, Elisabetta; Donati, Maria A.; Zara, Federico; Lisanti, Michael P.; Bruno, Claudio; Minetti, Carlo.

In: Laboratory Investigation, Vol. 88, No. 3, 03.2008, p. 275-283.

Research output: Contribution to journalArticle

Traverso, M, Gazzerro, E, Assereto, S, Sotgia, F, Biancheri, R, Stringara, S, Giberti, L, Pedemonte, M, Wang, X, Scapolan, S, Pasquini, E, Donati, MA, Zara, F, Lisanti, MP, Bruno, C & Minetti, C 2008, 'Caveolin-3 T78M and T78K missense mutations lead to different phenotypes in vivo and in vitro', Laboratory Investigation, vol. 88, no. 3, pp. 275-283. https://doi.org/10.1038/labinvest.3700713
Traverso, Monica ; Gazzerro, Elisabetta ; Assereto, Stefania ; Sotgia, Federica ; Biancheri, Roberta ; Stringara, Silvia ; Giberti, Laura ; Pedemonte, Marina ; Wang, Xiabo ; Scapolan, Sara ; Pasquini, Elisabetta ; Donati, Maria A. ; Zara, Federico ; Lisanti, Michael P. ; Bruno, Claudio ; Minetti, Carlo. / Caveolin-3 T78M and T78K missense mutations lead to different phenotypes in vivo and in vitro. In: Laboratory Investigation. 2008 ; Vol. 88, No. 3. pp. 275-283.
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abstract = "Caveolins are the principal protein components of caveolae, invaginations of the plasma membrane involved in cell signaling and trafficking. Caveolin-3 (Cav-3) is the muscle-specific isoform of the caveolin family and mutations in the CAV3 gene lead to a large group of neuromuscular disorders. In unrelated patients, we identified two distinct CAV3 mutations involving the same codon 78. Patient 1, affected by dilated cardiomyopathy and limb girdle muscular dystrophy (LGMD)-1C, shows an autosomal recessive mutation converting threonine to methionine (T78M). Patient 2, affected by isolated familiar hyperCKemia, shows an autosomal dominant mutation converting threonine to lysine (T78K). Cav-3 wild type (WT) and Cav-3 mutations were transiently transfected into Cos-7 cells. Cav-3 WT and Cav-3 T78M mutant localized at the plasma membrane, whereas Cav-3 T78K was retained in a perinuclear compartment. Cav-3 T78K expression was decreased by 87{\%} when compared with Cav-3 WT, whereas Cav-3 T78M protein levels were unchanged. To evaluate whether Cav-3 T78K and Cav-3 T78M mutants behaved with a dominant negative pattern, Cos-7 cells were cotransfected with green fluorescent protein (GFP)-Cav-3 WT in combination with either mutant or WT Cav-3. When cotransfected with Cav-3 WT or Cav-3 T78M, GFP-Cav-3 WT was localized at the plasma membrane, as expected. However, when cotransfected with Cav-3 T78K, GFP-Cav-3 WT was retained in a perinuclear compartment, and its protein levels were reduced by 60{\%}, suggesting a dominant negative action. Accordingly, Cav-3 protein levels in muscles from a biopsy of patient 2 (T78K mutation) were reduced by 80{\%}. In conclusion, CAV3 T78M and T78K mutations lead to distinct disorders showing different clinical features and inheritance, and displaying distinct phenotypes in vitro.",
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AU - Biancheri, Roberta

AU - Stringara, Silvia

AU - Giberti, Laura

AU - Pedemonte, Marina

AU - Wang, Xiabo

AU - Scapolan, Sara

AU - Pasquini, Elisabetta

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AU - Zara, Federico

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AU - Bruno, Claudio

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