Department of Medicine and Clinical Science, Division of Cardiology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan; Department of Pharmacology, University of California, Davis, CA, United States.
Department of Medicine and Clinical Science, Division of Cardiology, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi, Japan.
J Mol Cell Cardiol. 2018 Dec;125:87-97. doi: 10.1016/j.yjmcc.2018.10.011. Epub 2018 Oct 22.
In cardiac myocytes Calmodulin (CaM) bound to the ryanodine receptor (RyR2) constitutes a large pool of total myocyte CaM, but the CaM-RyR2 affinity is reduced in pathological conditions. Knock-in mice expressing RyR2 unable to bind CaM also developed hypertrophy and early death. However, it is unknown whether CaM released from this RyR2-bound pool participates in pathological cardiac hypertrophy. We found that angiotensin II (AngII) or phenylephrine (PE) both cause CaM to dissociate from the RyR2 and translocate to the nucleus. To test whether this nuclear CaM accumulation depends on CaM released from RyR2, we enhanced CaM-RyR2 binding affinity (with dantrolene), or caused CaM dissociation from RyR2 (using suramin). Dantrolene dramatically reduced AngII- and PE-induced nuclear CaM accumulation. Conversely, suramin enhanced nuclear CaM accumulation. This is consistent with nuclear CaM accumulation coming largely from the CaM-RyR2 pool. CaM lacks a nuclear localization signal (NLS), but G-protein coupled receptor kinase 5 (GRK5) binds CaM, has a NLS and translocates like CaM in response to AngII or PE. Suramin also promoted GRK5 nuclear import, and caused nuclear export of histone deacetylase 5 (HDAC5). Dantrolene prevented these effects. After 2-8 weeks of pressure overload (TAC) CaM binding to RyR2 was reduced, nuclear CaM and GRK5 were both elevated and there was enhanced nuclear export of HDAC5. Stress (acute AngII or TAC) causes CaM dissociation from RyR2 and translocation to the nucleus with GRK5 with parallel HDAC5 nuclear export. Thus CaM dissociation from RyR2 may be an important step in driving pathological hypertrophic gene transcription.
在心肌细胞中,与兰尼碱受体 2(RyR2)结合的钙调蛋白(CaM)构成了总心肌细胞 CaM 的一个大池,但在病理条件下,CaM-RyR2 亲和力降低。表达无法结合 CaM 的 RyR2 的基因敲入小鼠也发展为肥大和早期死亡。然而,尚不清楚从这种与 RyR2 结合的池释放的 CaM 是否参与病理性心肌肥大。我们发现血管紧张素 II(AngII)或苯肾上腺素(PE)均可使 CaM 从 RyR2 上解离并易位到核内。为了测试这种核内 CaM 聚集是否依赖于从 RyR2 释放的 CaM,我们增强了 CaM-RyR2 结合亲和力(使用丹曲林),或使 CaM 从 RyR2 上解离(使用苏拉明)。丹曲林显著减少了 AngII 和 PE 诱导的核内 CaM 聚集。相反,苏拉明增强了核内 CaM 聚集。这与核内 CaM 聚集主要来自 CaM-RyR2 池一致。CaM 缺乏核定位信号(NLS),但 G 蛋白偶联受体激酶 5(GRK5)与 CaM 结合,具有 NLS,并响应 AngII 或 PE 像 CaM 一样易位。苏拉明还促进了 GRK5 的核内导入,并导致组蛋白去乙酰化酶 5(HDAC5)的核输出。丹曲林阻止了这些作用。在压力超负荷(TAC)后 2-8 周,CaM 与 RyR2 的结合减少,核内 CaM 和 GRK5 均升高,HDAC5 的核输出增强。应激(急性 AngII 或 TAC)导致 CaM 从 RyR2 上解离并与 GRK5 一起易位到核内,同时伴有 HDAC5 的核输出。因此,CaM 从 RyR2 上的解离可能是驱动病理性肥大基因转录的重要步骤。