{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T16:03:53Z","timestamp":1753891433024,"version":"3.41.2"},"reference-count":32,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T00:00:00Z","timestamp":1734998400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:sec><jats:title>Aims<\/jats:title><jats:p>Mutations in the cardiac ryanodine receptor (RyR2) are associated with catecholaminergic polymorphic ventricular tachycardia (CPVT). This study investigates the underlying molecular mechanisms for CPVT mutations within the RyR2 N-terminus domain (NTD).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods and Results<\/jats:title><jats:p>We consulted the high-resolution RyR2 structure in both open and closed configuration to identify mutations G357S\/R407I and A77T, which lie within the NTD intra- and inter-subunit interface with the Core Solenoid (CSol), respectively. Their structural and functional roles were compared to R169L, a mutation that lies within the NTD-NTD inter-subunit interface. Using chemical cross-linking and co-immunoprecipitation assays, we show that R169L disrupts NTD tetramerization, while it does not alter the NTD-CSol interaction. Single cell Ca<jats:sup>2+<\/jats:sup> imaging revealed that R169L increases the number of spontaneous Ca<jats:sup>2+<\/jats:sup> transients and the proportion of oscillating cells, while reducing the Ca<jats:sup>2+<\/jats:sup> store content. G357S and R407I do not affect NTD tetramerization, but they also do not alter the NTD-CSol interaction. Functionally, RyR2<jats:sup>G357S<\/jats:sup>-expressing cells have Ca<jats:sup>2+<\/jats:sup> handling properties similar to RyR2<jats:sup>WT<\/jats:sup>. A77T enhances the NTD-CSol interaction, while it does not affect NTD tetramerization. Like R169L, A77T also increases the number of spontaneous Ca<jats:sup>2+<\/jats:sup> transients and the proportion of oscillating cells, and it reduces the Ca<jats:sup>2+<\/jats:sup> store content. However, unlike R169L that displays Ca<jats:sup>2+<\/jats:sup> transients of normal amplitude and shorter duration, Ca<jats:sup>2+<\/jats:sup> transients for A77T are of smaller amplitude and normal duration.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The NTD-CSol inter-subunit interface variant, A77T, produces a hyperactive channel by altering a different structure-function parameter to other CPVT mutations within the RyR2 NTD. Reduced NTD-NTD inter-subunit interaction and reinforced NTD inter-subunit interaction with CSol are distinct molecular mechanisms for gain-of-function RyR2 arrhythmogenic mutations.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmolb.2024.1505698","type":"journal-article","created":{"date-parts":[[2024,12,24]],"date-time":"2024-12-24T05:10:36Z","timestamp":1735017036000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Disparate molecular mechanisms in cardiac ryanodine receptor channelopathies"],"prefix":"10.3389","volume":"11","author":[{"given":"Yadan","family":"Zhang","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monika","family":"Seidel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Camille","family":"Rabesahala de Meritens","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Astrid","family":"Beckmann","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Syeda","family":"Ahmed","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melanie","family":"Hurtz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F. 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