• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Cell motility and the cytoskeleton >Targeted disruption of N-RAP gene function by RNA interference: a role for N-RAP in myofibril organization.
【24h】

Targeted disruption of N-RAP gene function by RNA interference: a role for N-RAP in myofibril organization.

机译：RNA干扰靶向破坏N-RAP基因功能：N-RAP在肌原纤维组织中的作用。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

N-RAP is a muscle-specific protein concentrated in myofibril precursors during sarcomere assembly and at intercalated disks in adult heart. We used RNA interference to achieve a targeted decrease in N-RAP transcript and protein levels in primary cultures of embryonic mouse cardiomyocytes. N-RAP transcript levels were decreased by approximately 70% within 2 days following transfection with N-RAP specific siRNA. N-RAP protein levels steadily decreased over several days, reaching approximately 50% of control levels within 6 days. N-RAP protein knockdown was associated with decreased myofibril assembly, as assessed by alpha-actinin organization into mature striations. Transcripts encoding N-RAP binding proteins associated with assembling or mature myofibrils, such as alpha-actinin, Krp1, and muscle LIM protein, were expressed at normal levels during N-RAP protein knockdown, and alpha-actinin and Krp-1 protein levels were also unchanged. Transcripts encoding muscle myosin heavy chain and nonmuscle myosin heavy chain IIB were also expressed at relatively normal levels. However, decreased N-RAP protein levels were associated with dramatic changes in the encoded myosin proteins, with muscle myosin heavy chain levels increasing and nonmuscle myosin heavy chain IIB decreasing. N-RAP transcript and protein levels recovered to normal by days 6 and 7, respectively, and the changes in myofibril organization and myosin heavy chain isoform levels were reversed. Our data indicate that we can achieve transient N-RAP protein knockdown using the RNA interference technique and that alpha-actinin organization into myofibrils in cardiomyocytes is closely linked to N-RAP protein levels. Finally, N-RAP protein levels regulate the balance between nonmuscle myosin IIB and muscle myosin by post-trancriptional mechanisms.
机译：N-RAP是一种肌特异性蛋白，在肌节组装过程中和成年心脏的插层中聚集在肌原纤维前体中。我们使用RNA干扰来实现胚胎小鼠心肌细胞原代培养中N-RAP转录和蛋白质水平的有针对性的降低。用N-RAP特异性siRNA转染后2天内，N-RAP转录水平降低了约70％。 N-RAP蛋白水平在几天内稳定下降，在6天内达到对照水平的约50％。 N-RAP蛋白敲低与肌原纤维组装减少有关，如α-肌动蛋白组织形成的成熟条纹所示。编码与组装或成熟的肌原纤维相关的N-RAP结合蛋白的转录本，例如α-肌动蛋白，Krp1和肌肉LIM蛋白，在N-RAP蛋白敲低期间以正常水平表达，而α-actinin和Krp-1蛋白水平为也不变。编码肌肌球蛋白重链和非肌球蛋白重链IIB的转录本也以相对正常的水平表达。但是，降低的N-RAP蛋白水平与编码的肌球蛋白蛋白的急剧变化有关，肌肉肌球蛋白重链水平增加而非肌球蛋白重链IIB下降。 N-RAP转录和蛋白质水平分别在第6天和第7天恢复正常，肌原纤维组织和肌球蛋白重链同工型水平的变化被逆转。我们的数据表明，我们可以使用RNA干扰技术实现瞬时N-RAP蛋白的敲低，并且心肌细胞中肌原纤维中的α-肌动蛋白组织与N-RAP蛋白水平密切相关。最后，N-RAP蛋白水平通过转录后机制调节非肌球蛋白IIB和肌球蛋白之间的平衡。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号