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Next-generation sequencing identifies transportin 3 as the causative gene for LGMD1F.

Torella A, Fanin M, Mutarelli M, Peterle E, Del Vecchio Blanco F, Rispoli R, Savarese M, Garofalo A, Piluso G, Morandi L, Ricci G, Siciliano G, Angelini C, Nigro V - PLoS ONE (2013)

Bottom Line: In addition, we identified an isolated case of LGMD with a new missense mutation in the same gene.We localized the mutant TNPO3 around the nucleus, but not inside.The involvement of gene related to the nuclear transport suggests a novel disease mechanism leading to muscular dystrophy.

View Article: PubMed Central - PubMed

Affiliation: TIGEM (Telethon Institute of Genetics and Medicine), Napoli, Italy.

ABSTRACT
Limb-girdle muscular dystrophies (LGMD) are genetically and clinically heterogeneous conditions. We investigated a large family with autosomal dominant transmission pattern, previously classified as LGMD1F and mapped to chromosome 7q32. Affected members are characterized by muscle weakness affecting earlier the pelvic girdle and the ileopsoas muscles. We sequenced the whole exome of four family members and identified a shared heterozygous frame-shift variant in the Transportin 3 (TNPO3) gene, encoding a member of the importin-β super-family. The TNPO3 gene is mapped within the LGMD1F critical interval and its 923-amino acid human gene product is also expressed in skeletal muscle. In addition, we identified an isolated case of LGMD with a new missense mutation in the same gene. We localized the mutant TNPO3 around the nucleus, but not inside. The involvement of gene related to the nuclear transport suggests a novel disease mechanism leading to muscular dystrophy.

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Indirect immunofluorescence analysis of the wt-hTNPO3 compared with delA p.X924C -hTNPO3.Following transient transfections, HeLa cells were incubated for 48 h with normal DMEM and detected by anti-HA immunofluorescence. Nuclei are stained with DAPI (blue). The endogenous protein is recognized using a rabbit monoclonal anti-TNPO3 antibody (green), while the transfected TNPO3 proteins were HA-tagged (red). a) An accumulation around the nucleus is usually observed using the mutant delA p.X924C -hTNPO3. b) The typical intranuclear staining pattern can be observed in cells transfected with wt-hTNPO3 (in red) or c) in non transfected HeLa cells.
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pone-0063536-g004: Indirect immunofluorescence analysis of the wt-hTNPO3 compared with delA p.X924C -hTNPO3.Following transient transfections, HeLa cells were incubated for 48 h with normal DMEM and detected by anti-HA immunofluorescence. Nuclei are stained with DAPI (blue). The endogenous protein is recognized using a rabbit monoclonal anti-TNPO3 antibody (green), while the transfected TNPO3 proteins were HA-tagged (red). a) An accumulation around the nucleus is usually observed using the mutant delA p.X924C -hTNPO3. b) The typical intranuclear staining pattern can be observed in cells transfected with wt-hTNPO3 (in red) or c) in non transfected HeLa cells.

Mentions: We generated a construct expressing the WT and del A p.X924C allele. HeLa cells were transfected with either the Wt or the mutant TNPO3. The transfected proteins were distinguished from the endogenous TNPO3 by adding a HA-tag. Figure 4 shows that the WT TNPO3 entered the nucleus, while the mutant was usually around the periphery of the nucleus.


Next-generation sequencing identifies transportin 3 as the causative gene for LGMD1F.

Torella A, Fanin M, Mutarelli M, Peterle E, Del Vecchio Blanco F, Rispoli R, Savarese M, Garofalo A, Piluso G, Morandi L, Ricci G, Siciliano G, Angelini C, Nigro V - PLoS ONE (2013)

Indirect immunofluorescence analysis of the wt-hTNPO3 compared with delA p.X924C -hTNPO3.Following transient transfections, HeLa cells were incubated for 48 h with normal DMEM and detected by anti-HA immunofluorescence. Nuclei are stained with DAPI (blue). The endogenous protein is recognized using a rabbit monoclonal anti-TNPO3 antibody (green), while the transfected TNPO3 proteins were HA-tagged (red). a) An accumulation around the nucleus is usually observed using the mutant delA p.X924C -hTNPO3. b) The typical intranuclear staining pattern can be observed in cells transfected with wt-hTNPO3 (in red) or c) in non transfected HeLa cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3646821&req=5

pone-0063536-g004: Indirect immunofluorescence analysis of the wt-hTNPO3 compared with delA p.X924C -hTNPO3.Following transient transfections, HeLa cells were incubated for 48 h with normal DMEM and detected by anti-HA immunofluorescence. Nuclei are stained with DAPI (blue). The endogenous protein is recognized using a rabbit monoclonal anti-TNPO3 antibody (green), while the transfected TNPO3 proteins were HA-tagged (red). a) An accumulation around the nucleus is usually observed using the mutant delA p.X924C -hTNPO3. b) The typical intranuclear staining pattern can be observed in cells transfected with wt-hTNPO3 (in red) or c) in non transfected HeLa cells.
Mentions: We generated a construct expressing the WT and del A p.X924C allele. HeLa cells were transfected with either the Wt or the mutant TNPO3. The transfected proteins were distinguished from the endogenous TNPO3 by adding a HA-tag. Figure 4 shows that the WT TNPO3 entered the nucleus, while the mutant was usually around the periphery of the nucleus.

Bottom Line: In addition, we identified an isolated case of LGMD with a new missense mutation in the same gene.We localized the mutant TNPO3 around the nucleus, but not inside.The involvement of gene related to the nuclear transport suggests a novel disease mechanism leading to muscular dystrophy.

View Article: PubMed Central - PubMed

Affiliation: TIGEM (Telethon Institute of Genetics and Medicine), Napoli, Italy.

ABSTRACT
Limb-girdle muscular dystrophies (LGMD) are genetically and clinically heterogeneous conditions. We investigated a large family with autosomal dominant transmission pattern, previously classified as LGMD1F and mapped to chromosome 7q32. Affected members are characterized by muscle weakness affecting earlier the pelvic girdle and the ileopsoas muscles. We sequenced the whole exome of four family members and identified a shared heterozygous frame-shift variant in the Transportin 3 (TNPO3) gene, encoding a member of the importin-β super-family. The TNPO3 gene is mapped within the LGMD1F critical interval and its 923-amino acid human gene product is also expressed in skeletal muscle. In addition, we identified an isolated case of LGMD with a new missense mutation in the same gene. We localized the mutant TNPO3 around the nucleus, but not inside. The involvement of gene related to the nuclear transport suggests a novel disease mechanism leading to muscular dystrophy.

Show MeSH
Related in: MedlinePlus