Disruption of the nuclear localization signal in RBM20 is causative in dilated cardiomyopathy.
Disruption of the nuclear localization signal in RBM20 is causative in dilated cardiomyopathy.
复制标题
DOI：
10.1172/jci.insight.170001
复制
发表时间：
2023-07-10
期刊：
影响因子：
8
通讯作者：
Guo W
中科院分区：
文献类型：
--
作者：
Zhang Y;Gregorich ZR;Wang Y;Braz CU;Zhang J;Liu Y;Liu P;Shen J;Aori N;Hacker TA;Granzier H;Guo W
携带RNA结合基序20（RBM20）中的基因突变的人类患者发展为临床上侵袭性扩张型心肌病（DCM）。基因突变敲入（KI）动物模型表明，富含丝氨酸（RS）结构域的功能改变对严重DCM至关重要。为了验证这一假设，我们建立了RS结构域缺失小鼠模型（Rbm20ΔRS）。我们发现Rbm20ΔRS小鼠表现为DCM，伴有RBM20靶转录物的错误剪接。我们发现RBM20错误定位于Rbm20ΔRS小鼠心脏的肌浆中，并形成类似于突变KI动物中检测到的RBM20颗粒。相比之下，缺乏RNA识别基序的小鼠显示主要RBM20靶基因的类似错误剪接，但没有发展DCM或显示RBM20颗粒形成。使用免疫细胞化学染色的体外研究，我们证明，只有DCM相关的RS结构域的突变促进RBM20核质运输和促进颗粒组装。此外，我们在RBM20的RS结构域内定义了核心核定位信号（NLS）。RS结构域中磷酸化位点的突变分析表明，这种修饰可能与RBM20核质转运有关。总的来说，我们的研究结果表明，RS结构域介导的核定位的破坏是由NLS突变引起的严重DCM的关键。
Human patients carrying genetic mutations in RNA binding motif 20 (RBM20) develop a clinically aggressive dilated cardiomyopathy (DCM). Genetic mutation knockin (KI) animal models imply that altered function of the arginine-serine-rich (RS) domain is crucial for severe DCM. To test this hypothesis, we generated an RS domain deletion mouse model (Rbm20ΔRS). We showed that Rbm20ΔRS mice manifested DCM with mis-splicing of RBM20 target transcripts. We found that RBM20 was mis-localized to the sarcoplasm in Rbm20ΔRS mouse hearts and formed RBM20 granules similar to those detected in mutation KI animals. In contrast, mice lacking the RNA recognition motif showed similar mis-splicing of major RBM20 target genes but did not develop DCM or exhibit RBM20 granule formation. Using in vitro studies with immunocytochemical staining, we demonstrated that only DCM-associated mutations in the RS domain facilitated RBM20 nucleocytoplasmic transport and promoted granule assembly. Further, we defined the core nuclear localization signal (NLS) within the RS domain of RBM20. Mutation analysis of phosphorylation sites in the RS domain suggested that this modification may be dispensable for RBM20 nucleocytoplasmic transport. Collectively, our findings revealed that disruption of RS domain–mediated nuclear localization is crucial for severe DCM caused by NLS mutations.
登录
查看更多内容
影响因子：
4.6
作者：
Ihara K;Sasano T;Hiraoka Y;Togo-Ohno M;Soejima Y;Sawabe M;Tsuchiya M;Ogawa H;Furukawa T;Kuroyanagi H
通讯作者：
Kuroyanagi H
影响因子：
16.6
作者：
Fenix AM;Miyaoka Y;Bertero A;Blue SM;Spindler MJ;Tan KKB;Perez-Bermejo JA;Chan AH;Mayerl SJ;Nguyen TD;Russell CR;Lizarraga PP;Truong A;So PL;Kulkarni A;Chetal K;Sathe S;Sniadecki NJ;Yeo GW;Murry CE;Conklin BR;Salomonis N
通讯作者：
Salomonis N
影响因子：
3.5
作者：
Gaertner A;Bloebaum J;Brodehl A;Klauke B;Sielemann K;Kassner A;Fox H;Morshuis M;Tiesmeier J;Schulz U;Knoell R;Gummert J;Milting H
通讯作者：
Milting H
DOI：
10.1073/pnas.181354098
发表时间：
2001-08-28
影响因子：
11.1
作者：
Lai, MC;Lin, RI;Tarn, WY
通讯作者：
Tarn, WY
影响因子：
10.8
作者：
Beqqali, Abdelaziz;Bollen, Ilse A. E.;Pinto, Yigal M.
通讯作者：
Pinto, Yigal M.