Journal article
A novel ANO5 splicing variant in a LGMD2L patient leads to production of a truncated aggregation‐prone Ano5 peptide
The journal of pathology. Clinical research, Vol.4(2), pp.135-145
04/2018
DOI: 10.1002/cjp2.92
PMCID: PMC5903698
PMID: 29665321
Abstract
Mutations in
ANO5
cause several human diseases including gnathodiaphyseal dysplasia 1 (GDD1), limb‐girdle muscular dystrophy 2L (LGMD2L), and Miyoshi myopathy 3 (MMD3). Previous work showed that complete genetic disruption of
Ano5
in mice did not recapitulate human muscular dystrophy, while residual expression of mutant
Ano5
in a gene trapped mouse developed muscular dystrophy with defective membrane repair. This suggests that truncated Ano5 expression may be pathogenic. Here, we screened a panel of commercial anti‐Ano5 antibodies using a recombinant adenovirus expressing human Ano5 with FLAG and YFP at the N‐ and C‐terminus, respectively. The monoclonal antibody (mAb) N421A/85 was found to specifically detect human Ano5 by immunoblotting and immunofluorescence staining. The antigen epitope was mapped to a region of 28 residues within the N‐terminus. Immunofluorescence staining of muscle cryosections from healthy control subjects showed that Ano5 is localized at the sarcoplasmic reticulum. The muscle biopsy from a LGMD2L patient homozygous for the c.191dupA mutation showed no Ano5 signal, confirming the specificity of the N421A/85 antibody. Surprisingly, strong Ano5 signal was detected in a patient with compound heterozygous mutations (c.191dupA and a novel splice donor site variant c.363 + 4A > G at the exon 6–intron 6 junction). Interestingly, insertion of the mutant intron 6, but not the wild‐type intron 6, into human
ANO5
cDNA resulted in a major transcript that carried the first 158‐bp of intron 6. Transfection of the construct encoding the first 121 amino acids into C2C12 cells resulted in protein aggregate formation, suggesting that aggregate‐forming Ano5 peptide may contribute to the pathogenesis of muscular dystrophy.
Details
- Title: Subtitle
- A novel ANO5 splicing variant in a LGMD2L patient leads to production of a truncated aggregation‐prone Ano5 peptide
- Creators
- Jing Xu - The Ohio State University Wexner Medical CenterLi Xu - The Ohio State University Wexner Medical CenterYeh S Lau - The Ohio State University Wexner Medical CenterYandi Gao - The Ohio State University Wexner Medical CenterSteven A Moore - University of IowaRenzhi Han - The Ohio State University Wexner Medical Center
- Resource Type
- Journal article
- Publication Details
- The journal of pathology. Clinical research, Vol.4(2), pp.135-145
- Publisher
- John Wiley and Sons Inc; Hoboken
- DOI
- 10.1002/cjp2.92
- PMID
- 29665321
- PMCID
- PMC5903698
- ISSN
- 2056-4538
- eISSN
- 2056-4538
- Grant note
- Iowa Wellstone Muscular Dystrophy Cooperative Research Center HL116546; AR064241; U54; NS053672 / US National Institutes of Health
- Alternative title
- A novel ANO5 splicing variant in LGMD2L
- Language
- English
- Date published
- 04/2018
- Academic Unit
- Pathology
- Record Identifier
- 9984047622302771
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