Limits...
A Haplotype of Two Novel Polymorphisms in δ-Sarcoglycan Gene Increases Risk of Dilated Cardiomyopathy in Mongoloid Population.

Chen J, Jin Y, Wang H, Wei S, Chen D, Ying L, Zhou Q, Li G, Li J, Gao J, Kato N, Hu W, Li Y, Wang Y - PLoS ONE (2015)

Bottom Line: C.-100~-110 resulted in a decrease of δ-SG promoter activity to 64±3% of the control level (p<0.01).Both co-immunoprecipitation and in vitro protein pull-down assays demonstrated that δ-SG-283R interacts normally to β- and γ-SG, but significantly decreased localization of β/δ/γ-SG on the plasma membrane.In conclusion, haplotype -_G composed of c.-100~-110 and A848G confers higher susceptibility to DCM in the Mongoloid population.

View Article: PubMed Central - PubMed

Affiliation: Molecular Cardiology Research Laboratory, Department of Cardiology, Affiliated Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.

ABSTRACT
The role of genetic abnormality of δ-sarcoglycan (δ-SG) gene in dilated (DCM) and hypertrophied (HCM) cardiomyopathy patients is still unfolding. In this study we first defined the promoter region and then searched for polymorphisms/mutations among the promoter, 5'-untranslated region, and the encoding exons in δ-SG gene in 104 Chinese patients with DCM, 145 with HCM, and 790 normal controls. Two novel polymorphisms were found, an 11 base-pair (bp) deletion (c.-100~-110; -) in the promoter region and a missense polymorphism of A848G resulting in p.Q283R in the highly conserved C-terminus. The prevalence of homozygous genotype -/- of c.-100~-110 was slightly higher in DCM (14.42%) and HCM patients (14.48%), as compared with normal controls (11.01%). The prevalence of genotype of 848A/G was significantly higher in DCM (6.73%; OR = 9.43; p = 0.0002), but not in HCM patients (1.38%; OR = 1.37; p = 0.62), as compared with controls (0.76%). Haplotype -_G consisting c.-100~-110 and A848G was associated with increased risk of DCM (OR = 17.27; 95%CI = 3.19-93.56; p = 0.001) but not associated with HCM (OR = 1.90; 95%CI = 0.38-9.55; p = 0.44). Co-occurrence of the genotypes -/- of c.-100~-110 and 848A/G was found in 5 patients with DCM (4.81%; OR = 39.85; p = 0.0001), none of HCM patients, and only 1 of the controls (0.13%). Both polymorphisms were also found in the Japanese population, but not in the Africans and Caucasians. C.-100~-110 resulted in a decrease of δ-SG promoter activity to 64±3% of the control level (p<0.01). Both co-immunoprecipitation and in vitro protein pull-down assays demonstrated that δ-SG-283R interacts normally to β- and γ-SG, but significantly decreased localization of β/δ/γ-SG on the plasma membrane. In conclusion, haplotype -_G composed of c.-100~-110 and A848G confers higher susceptibility to DCM in the Mongoloid population.

Show MeSH

Related in: MedlinePlus

Genotypic analysis of the promoter-containing F-8.Fig 2A shows the normal genomic sequence of -90~-120 within the 5’-UTR. Fig 2B and 2C demonstrate the location (*) of the heterozygous deletion deduced from overlapped waves respectively in forward (B) and in reverse (C) directions (arrows). Fig 2D shows a homozygous c.-100~-110. Fig 2E shows losing of a Hind III site due to the c.-100~-110. Fig 2F illustrates differential cutting patterns of normal (N; double bands of 237 and 577bp), heterozygous (He; triple bands of 237, 577, and 803bp), and homozygous (Ho; single band of 803bp) c.-100~-110 in F-8 in Orientation, African, and Caucasian populations. The cut samples were separated and visualized on 2% agarose ethidium bromide gel.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4697846&req=5

pone.0145602.g002: Genotypic analysis of the promoter-containing F-8.Fig 2A shows the normal genomic sequence of -90~-120 within the 5’-UTR. Fig 2B and 2C demonstrate the location (*) of the heterozygous deletion deduced from overlapped waves respectively in forward (B) and in reverse (C) directions (arrows). Fig 2D shows a homozygous c.-100~-110. Fig 2E shows losing of a Hind III site due to the c.-100~-110. Fig 2F illustrates differential cutting patterns of normal (N; double bands of 237 and 577bp), heterozygous (He; triple bands of 237, 577, and 803bp), and homozygous (Ho; single band of 803bp) c.-100~-110 in F-8 in Orientation, African, and Caucasian populations. The cut samples were separated and visualized on 2% agarose ethidium bromide gel.

Mentions: Next, we searched for SNP/mutations in the 814bp promoter region in CM patients by direct sequencing. In some samples, compared to the WT (Fig 2A), overlapped spikes always started from a certain area in both forward (Fig 2B) and reverse (Fig 2C) sequencing directions. These overlapping spikes disappeared when we sequenced a sample from a patient carrying an 11bp-deletion in both alleles. This indicated that the overlapped sequencing spikes was due to a heterozygous 11bp-deletion (c.-100~-110; Chrom-5: 155,753,657–155,753,667; Fig 2D). C.-100~-110 locates just downstream of the E-2 and this AT-rich sequence (TGTAATAAGCT) itself likely constitutes the third putative MEF2C binding motif (MEF2C-3). The c.-100~-110 abolished unique Hind III site (Fig 2E), which was used as an easy identification in mass genotyping (Fig 2F).


A Haplotype of Two Novel Polymorphisms in δ-Sarcoglycan Gene Increases Risk of Dilated Cardiomyopathy in Mongoloid Population.

Chen J, Jin Y, Wang H, Wei S, Chen D, Ying L, Zhou Q, Li G, Li J, Gao J, Kato N, Hu W, Li Y, Wang Y - PLoS ONE (2015)

Genotypic analysis of the promoter-containing F-8.Fig 2A shows the normal genomic sequence of -90~-120 within the 5’-UTR. Fig 2B and 2C demonstrate the location (*) of the heterozygous deletion deduced from overlapped waves respectively in forward (B) and in reverse (C) directions (arrows). Fig 2D shows a homozygous c.-100~-110. Fig 2E shows losing of a Hind III site due to the c.-100~-110. Fig 2F illustrates differential cutting patterns of normal (N; double bands of 237 and 577bp), heterozygous (He; triple bands of 237, 577, and 803bp), and homozygous (Ho; single band of 803bp) c.-100~-110 in F-8 in Orientation, African, and Caucasian populations. The cut samples were separated and visualized on 2% agarose ethidium bromide gel.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4697846&req=5

pone.0145602.g002: Genotypic analysis of the promoter-containing F-8.Fig 2A shows the normal genomic sequence of -90~-120 within the 5’-UTR. Fig 2B and 2C demonstrate the location (*) of the heterozygous deletion deduced from overlapped waves respectively in forward (B) and in reverse (C) directions (arrows). Fig 2D shows a homozygous c.-100~-110. Fig 2E shows losing of a Hind III site due to the c.-100~-110. Fig 2F illustrates differential cutting patterns of normal (N; double bands of 237 and 577bp), heterozygous (He; triple bands of 237, 577, and 803bp), and homozygous (Ho; single band of 803bp) c.-100~-110 in F-8 in Orientation, African, and Caucasian populations. The cut samples were separated and visualized on 2% agarose ethidium bromide gel.
Mentions: Next, we searched for SNP/mutations in the 814bp promoter region in CM patients by direct sequencing. In some samples, compared to the WT (Fig 2A), overlapped spikes always started from a certain area in both forward (Fig 2B) and reverse (Fig 2C) sequencing directions. These overlapping spikes disappeared when we sequenced a sample from a patient carrying an 11bp-deletion in both alleles. This indicated that the overlapped sequencing spikes was due to a heterozygous 11bp-deletion (c.-100~-110; Chrom-5: 155,753,657–155,753,667; Fig 2D). C.-100~-110 locates just downstream of the E-2 and this AT-rich sequence (TGTAATAAGCT) itself likely constitutes the third putative MEF2C binding motif (MEF2C-3). The c.-100~-110 abolished unique Hind III site (Fig 2E), which was used as an easy identification in mass genotyping (Fig 2F).

Bottom Line: C.-100~-110 resulted in a decrease of δ-SG promoter activity to 64±3% of the control level (p<0.01).Both co-immunoprecipitation and in vitro protein pull-down assays demonstrated that δ-SG-283R interacts normally to β- and γ-SG, but significantly decreased localization of β/δ/γ-SG on the plasma membrane.In conclusion, haplotype -_G composed of c.-100~-110 and A848G confers higher susceptibility to DCM in the Mongoloid population.

View Article: PubMed Central - PubMed

Affiliation: Molecular Cardiology Research Laboratory, Department of Cardiology, Affiliated Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.

ABSTRACT
The role of genetic abnormality of δ-sarcoglycan (δ-SG) gene in dilated (DCM) and hypertrophied (HCM) cardiomyopathy patients is still unfolding. In this study we first defined the promoter region and then searched for polymorphisms/mutations among the promoter, 5'-untranslated region, and the encoding exons in δ-SG gene in 104 Chinese patients with DCM, 145 with HCM, and 790 normal controls. Two novel polymorphisms were found, an 11 base-pair (bp) deletion (c.-100~-110; -) in the promoter region and a missense polymorphism of A848G resulting in p.Q283R in the highly conserved C-terminus. The prevalence of homozygous genotype -/- of c.-100~-110 was slightly higher in DCM (14.42%) and HCM patients (14.48%), as compared with normal controls (11.01%). The prevalence of genotype of 848A/G was significantly higher in DCM (6.73%; OR = 9.43; p = 0.0002), but not in HCM patients (1.38%; OR = 1.37; p = 0.62), as compared with controls (0.76%). Haplotype -_G consisting c.-100~-110 and A848G was associated with increased risk of DCM (OR = 17.27; 95%CI = 3.19-93.56; p = 0.001) but not associated with HCM (OR = 1.90; 95%CI = 0.38-9.55; p = 0.44). Co-occurrence of the genotypes -/- of c.-100~-110 and 848A/G was found in 5 patients with DCM (4.81%; OR = 39.85; p = 0.0001), none of HCM patients, and only 1 of the controls (0.13%). Both polymorphisms were also found in the Japanese population, but not in the Africans and Caucasians. C.-100~-110 resulted in a decrease of δ-SG promoter activity to 64±3% of the control level (p<0.01). Both co-immunoprecipitation and in vitro protein pull-down assays demonstrated that δ-SG-283R interacts normally to β- and γ-SG, but significantly decreased localization of β/δ/γ-SG on the plasma membrane. In conclusion, haplotype -_G composed of c.-100~-110 and A848G confers higher susceptibility to DCM in the Mongoloid population.

Show MeSH
Related in: MedlinePlus