CAPON Gene Structure, Isoforms, and Protein Functional Domains

<div><p>CAPON full-length transcript and protein illustrations are based on NCBI reference sequences.</p> <p>(A) Genomic organization of <i>CAPON</i>. Exons are represented by numbered boxes; 5′ and 3′ UTR are represented by half-height boxes.</p> <p>(B) <i>CAPON</i> transcripts. 5′ and 3′ UTR are represented by yellow shaded boxes, exons by white boxes.</p> <p>(C) CAPON proteins. The phosphotyrosine-binding domain (PTB) is shaded green, and the PDZ-binding domain is shaded blue.</p></div

ACTB (Beta-Actin)–Normalized <i>CAPON</i> mRNA Short-Form Expression by Genotype

<p>Expression levels are least squares means. Individuals from all three diagnostic classifications are included, grouped only by genotype. Mean values per genotype for each SNP are plotted with 95% confidence intervals. The number of individuals per genotype is indicated within each bar. SNP alleles are given for forward strand sequence. All three SNPs exhibit significantly (<i>p</i> < 0.05) different levels of <i>CAPON</i> expression by genotype, with a dominant effect. Higher levels of <i>CAPON</i> are seen in individuals with one or two copies of alleles previously identified as associated with schizophrenia (T, rs1415263; C, rs4145621; and C, rs2661818). The mean (95% confidence interval lower bound, upper bound) for the three genotypes for each SNP are as follows. For rs1415263: 1.54 (1.29, 1.80) for CC; 2.07 (1.81, 2.34) for CT; and 1.83 (1.36, 2.29) for TT. For rs4145621: 1.51 (1.25, 1.78) for TT; 2.01 (1.74, 2.27) for TC; and 2.01 (1.61, 2.41) for CC. For rs2661818: 1.49 (1.21, 1.76) for GG; 1.96 (1.70, 2.23) for CG; and 2.04 (1.68, 2.41) for CC.</p

ACTB (Beta-Actin)–-Normalized <i>CAPON</i> mRNA Full-Length Expression by Diagnosis

<p>Expression levels are least squares means. Mean values per category are plotted with 95% confidence intervals. The number of individuals per sample is indicated within each bar. Level of expression does not differ significantly by diagnostic group. The mean (95% confidence interval lower bound, upper bound) for the control, schizophrenia, and bipolar groups are 1.28 (1.12, 1.45), 1.33 (1.17, 1.49), and 1.16 (0.99, 1.32), respectively.</p

ACTB (Beta-Actin)–Normalized <i>CAPON</i> mRNA Short-Form Expression by Diagnosis

<p>Expression levels are least squares means. Mean values per category are plotted with 95% confidence intervals. The number of individuals per sample is indicated within each bar. Expression is significantly higher in patients with schizophrenia (<i>p</i> = 0.0013) and bipolar (<i>p</i> = 0.0009) as compared to controls. The mean (95% confidence interval lower bound, upper bound) for the control, schizophrenia, and bipolar groups are 1.34 (1.05, 1.62), 2.02 (1.73, 2.30), and 2.05 (1.77, 2.34), respectively.</p

Western Blot of CAPON Protein Isoforms in DLPFC from Normal Control Individuals

<p>Tissue from Brodmann's area 46 (DLPFC) from five individuals was homogenized in TEE. Proteins were resolved by SDS-PAGE and transferred to PVDF membrane. Blots were probed with rabbit polyclonal antibodies to CAPON and actin, and proteins were detected using chemiluminescence. Band intensities for CAPON-L, CAPON-S, and CAPON-S + CAPON-S′ were calculated and normalized to the intensities of the corresponding actin bands. Untransfected COS-7 cells expressing CAPON-L and CAPON-S′ (COS-7/NT) and COS-7 cells expressing recombinant CAPON-S (COS-7/CAPON-S) and were used as controls for these proteins. CAPON-L appears to include multiple bands, possibly due to phosphorylation.</p

Validation of a microRNA target site polymorphism in <i>H3F3B</i> that is potentially associated with a broad schizophrenia phenotype

<div><p>Despite much progress, few genetic findings for schizophrenia have been assessed by functional validation experiments at the molecular level. We previously reported evidence for genetic linkage of broadly defined schizophrenia to chromosome 17q25 in a sample of 24 multiplex families. 2,002 SNPs under this linkage peak were analyzed for evidence of linkage disequilibrium using the posterior probability of linkage (PPL) framework. SNP rs1060120 produced the strongest evidence for association, with a PPLD|L score of 0.21. This SNP is located within the 3'UTR of the histone gene <i>H3F3B</i> and colocalizes with potential gene target miR-616. A custom miRNA target prediction program predicted that the binding of miR-616 to <i>H3F3B</i> transcripts would be altered by the allelic variants of rs1060120. We used dual luciferase assays to experimentally validate this interaction. The rs1060120 A allele significantly reduced luciferase expression, indicating a stronger interaction with miR-616 than the G allele (p = 0.000412). These results provide functional validation that this SNP could alter schizophrenia epigenetic mechanisms thereby contributing to schizophrenia-related disease risk.</p></div

Linkage disequilibrium between 1,544 SNPs and broad schizophrenia spectrum phenotype.

<p>PPLD|L values for 1,544 SNPs, including five MirSNPs (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0194233#pone.0194233.t001" target="_blank">Table 1</a>), from chr17: 74,684,647 to 83,257,441 (GRCh38), were calculated using KELVIN v2.4.0 and plotted vs physical distance. The MirSNP rs1060120 in <i>H3F3B</i> produced a PPLD|L of 0.21, notably higher than the remaining SNPs.</p

Predicted MirSNPs from the chromosome 17q25 region of association/linkage to broadly defined schizophrenia.

<p>Predicted MirSNPs from the chromosome 17q25 region of association/linkage to broadly defined schizophrenia.</p

Regional Association Plots for Age at Natural Menopause in African American women in the PAGE Study.

<p>Locus Zoom plots for selected gene regions in age at natural menopause analysis. Vertical axis is the –log₁₀ of the p-value, the horizontal axis is the chromosomal position. Each dot represents a SNP tested for association with age at natural menopause in 1,860 African American women from the PAGE Study. Linkage disequilibrium between the most significant SNP, listed at the top of each plot, and the other SNPs in the plot is shown by the r² legend in each plot. (A) Locus Zoom plot for the <i>APOE</i> region, with rs78916952 the most significant SNP in the region. (B) Locus Zoom plot for the <i>MCM8</i> region; rs237688 is the most significant SNP in the plot region. (C) <i>FSHB</i> region Locus Zoom plot; rs605765 is the most significant SNP in the plot region. (D) Locus Zoom plot of the <i>BRSK1</i> region with rs11672111 as the most significant SNP in the plot region.</p

ANM Discovery–SNPs associated with age at natural menopause (ANM) in African American women from the PAGE Study.

<p>Tests of association at p≤1E-04 from single SNP linear regressions adjusted for study site and principal components in 1,860 African American women from the PAGE Study are shown. For each significant test of association, the chromosome, rs number, nearest gene, location, coded allele, beta, standard error (SE), and p-value are given. Genes listed are nearest genes to the SNP as measured from the transcription start site for upstream SNPs or the transcription stop site for downstream SNPs. Abbreviations: CAF, coded allele frequency.</p