Dissection of Z-disc myopalladin gene network involved in the development of restrictive cardiomyopathy using system genetics approach

Figure 1 Rank-ordered expression of Mypn in the heart across the 40 BXD strains and their parental strains. The X-axis denotes the strain name while the Y-axis denotes the mean expression given in a LOG2 scale.

Figure 2 Genetic mapping of Mypn expression in the heart of BXD mice. The interval mapping at the transcript level identified 4 suggestive eQTLs at chromosome 1, 5, 12, and X respectively (A). The interval mapping for the first principal component of exon 6, 12, and 17 showed a significant eQTL (genome-wide P < 0.05) at Chr 1 (B). The interval mapping for the first principal component of exons 7, 14, 18, and 19 showed a suggestive eQTL at Chr 1 and a significant eQTL (genome-wide P < 0.05) at Chr 5 (Figure 2C). The left Y-axis provides LRS score in blue and right Y-axis provides the additive effect in green. The red and green lines show the effect of the D or B allele on trait values, respectively. The upper X-axis shows location by chromosome and the lower X-axis shows location in megabases. The two horizontal lines across the plot make the threshold for genome-wide significant (P < 0.05, red or upper line) and suggestive (P < 0.63, grey or lower line) thresholds. eQTL: Expression quantitative trait locus; LRS: Likelihood ratio statistic.

Figure 3 Mypn gene network graph created using Gene-set Cohesion Analysis Tool described in the methods. Gene symbols are located at nodes in circles and lines interconnecting the nodes are based on literature correlation.

Figure 4 Two-dimensional gel electrophoresis of heart lysates from 12-wk-old mice. Comparative proteomics analysis revealed 10 non-redundant proteins in KI (heterozygote mutant) vs WT controls (A), 8 non-redundant proteins in KO (homozygote mutant) vs WT mouse hearts (B); 19 non-redundant protein changes in KO vs KI (C). Arrows indicate differential phisphorylation of proteins in WT vs KO and KI vs KO mice hearts (B and C, respectively).

Figure 5 Structure of Mypn gene and functional domain of the protein. The N-terminal domain containing two immunoglobulin (Ig) and coiled-coil domains binds to cardiac ankyrin repeat protein (Carp/Ankrd1), the negative regulator of muscle gene expression. The rod domain contains proline rich domain with phosphorylation residues and binds to the SH3-domain of nebulin/nebulette at the Z-discs. The C-terminal domain containing 3 Ig domains binds to α-actinin at the Z-discs.