FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Ubiquitination defect in cells expressing mutant FBXO32. a Co-immunopricipitation analysis. HEK293 cells were transfected with the indicated plasmids and immunoblot analysis was performed from total cell lysates using a specific anti-ubiquitin antibody. FBXO32 expression is shown as well as GAPDH. The blot is representative of three independent experiments. b Immunoblot analysis of the ubiquitination in cardiomyocytes. Cells were transfected with the Flag-FBXO32-WT or Flag-FBXO32-Mutant and whole cell extracts were analyzed by immunoblotting using the indicated antibodies. (TIF 1928 kb

Additional file 2: Figure S1. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Echocardiographic images of patients from the studied family. Echocardiography performed on two of the affected siblings index patients IV.5 (a) and IV.7 (b) prior to heart transplantation. M-mode echo and parasternal long-axis view showing dilatation of the left ventricle and of the left atria with severe hypokinesia. (TIF 5479 kb

Additional file 7: Figure S5. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Expression of the FBXO32 substrates calcineurin A and IkB-ι in FDC and IDC. Immunoblot analysis showing calcineurin A and IkB-ι protein expression in control hearts, in the heart of patients IV.5 and IV.7 carrying the FBXO32 variant, in a cardiomyopathic heart from another family (FDC2) and in idiopathic dilated hearts (IDC). FBXO32 and GAPDH are also shown. (TIF 3699 kb

Additional file 1: Table S1. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Clinical characteristics of the studied family. (DOCX 82 kb

Additional file 5: Figure S3. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Mutation in FBXO32 abrogates binding to SKP1 within the SCF complex. a Schematic representation of the SCF complex. The FBXO32 mutation represented as a black triangle is within the F-Box domain which normally mediates binding to SKP1. b Immunoprecipitation showing reduced interaction of mutant FBXO32 with CUL1 after co-transfection of HEK293 cells with WT-FBXO32 or mutant-FBXO32 and CUL1, ROC1, and SKP1. c Quantitative analysis of (b) from three independent experiments. (TIF 2185 kb

Additional file 3: Figure S2. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Autozygome analysis in the studied family. Autozygosity mapping was performed using AgileMultideogram. The result is showing the single shared ROH (runs of homozygosity) on chromosome 8 between the four affected members (dark blue). (TIF 4051 kb

Additional file 4: Table S2. of FBXO32, encoding a member of the SCF complex, is mutated in dilated cardiomyopathy

Coverage and sequencing depth of the exome data. (DOCX 17 kb