Therefore, we aimed at identifying other molecules potentially responding to the SdhD deletion

imilar extent in TG100 115 web wild-type and mutant hearts in late stage severe cardiac hypertrophy. Inhibition of CaMKII did not significantly alter the development of cardiac hypertrophy of Ryr2ADA/ADA mice. CaMKII inhibition resulted in a significant reduction of PLN phosphorylation at Thr-17 in Ryr2+/+ and Ryr2ADA/ADA hearts expressing AC3-I. We generated and characterized a second mutant mouse model with a single amino acid substitution in the CaM binding domain. In contrast to Ryr2ADA/ADA mice, Ryr2D/D mice showed loss of CaM inhibition at diastolic but not systolic Ca2+, had a normal lifespan, and showed only modest changes in heart size and function. This suggested that loss of CaM regulation in Ryr2D/D mice does not have a major impact as in Ryr2ADA/ADA mice, implying that CaM inhibition of RyR2 at systolic Ca2+ is necessary for normal cardiac growth and function. This was an unexpected finding because a “leaky”RyR2 at diastolic Ca2+, caused by phosphorylation of RyR2 at Ser2815, has been implicated in cardiomyopathies. Our results suggest that CaMKII did not significantly alter the development of cardiac hypertrophy of Ryr2ADA/ADA mice at either diastolic or systolic Ca2+. Protein kinase D, a member of the CaMK superfamily, has been reported to be inhibited by AC3-I. While a direct interaction with RyR2 has not been established, PKD, like CaMKII, phosphorylates class II histone deacetylases, which triggers their nuclear export, thereby regulating transcription and promoting cardiac remodeling. Immunoblots did not show significant differences in total PKD, pPKD-S744/S748 and pPKD-S916 protein levels between 10-day old wild-type and mutant mice expressing AC3 control or inhibitory peptides. However, phosphorylation at these sites does not necessarily reflect PDK activity in mutant and wild-type hearts. PKD binds diacylglycerol, which is formed by a variety of cellular stimuli, inducing cellular redistribution of the enzyme, and potentially altering PKD activity. Therefore, physiological relevance of in vitro measurements is uncertain, using the isolated form of the enzyme. To our knowledge there are no unique targets for PKD that could have been used to assess in vivo PKD activity. In conclusion, the results PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19659572 suggest that CaMKII does not have an essential role in the rapid development of cardiac hypertrophy and early death of mutant mice impaired in CaM regulation of RyR2 at diastolic and systolic Ca2+ concentrations. Although RyR2 mutations that impair CaM regulation in cardiac pathologies have not yet been identified, our studies should help to understand the role of RyR2 regulation by CaM and CaMKII in cardiac hypertrophy and heart failure in humans. Ryr2ADA/ADA Mice and AC3 Peptides The InterFeron Inducible TransMembrane protein family of viral restriction factors was defined in 2009 in a screen for host-modifying proteins of influenza A virus infection. Initially identified over 30 years ago, and named 927, 1-8D and 1-8U , these proteins received little attention until human IFITM3 depletion was found to enhance IAV infection in cell culture assays. Conversely, overexpression of IFITM3, or the closely related IFITM1 and IFITM2, could inhibit IAV replication. Studies in mice and humans have suggested that IFITM3, at least, also protects against IAV infection in vivo. However, the IFITM proteins are not IAV-specific and, in cell culture at least, have broad-spectrum antiviral activity, though in some systems they may enhance cellular infe