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Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
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During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
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在PLEKHM2 mRNA解码过程中编程的核糖体框架转移会产生一个构成性活跃的蛋白形,支持心肌功能.

Gary Loughran1, Raffaella De Pace2, Ningyu Ding3

  • 1School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland.

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概括

科学家们在人类基因PLEKHM2中发现了编程的核糖体框架转移,这是核基因中罕见的事件. 这种框架转移对于正常的心脏细胞功能至关重要.

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科学领域:

  • 分子生物学分子生物学
  • 遗传学 是一个遗传学.
  • 细胞生物学 细胞生物学

背景情况:

  • 编程核糖体框架转移是一种已知的机制,主要在病毒中,用于改变蛋白质合成.
  • 核编码基因中保存的框架转移事件很少见,并且经常引起争论.
  • 基因PLEKHM2在细胞过程中发挥作用,但其完整的调节机制尚未完全理解.

研究的目的:

  • 研究人类基因PLEKHM2.2.中的编程核糖体框架转移现象.
  • 描述这种移事件的机制和功能意义.
  • 确定框架转移在PLEKHM2的功能和细胞定位中的作用.

主要方法:

  • 在PLEKHM2.2翻译过程中分析mRNA序列和核糖体行为.
  • 位点定向的突变发生,以识别移信号.
  • 使用显微镜进行细胞局部化研究.
  • 在PLEKHM2-Knockout心肌细胞中的功能测试.

主要成果:

  • 在人类PLEKHM2基因中,在保存的UCC_UUU_CGG序列中确定了一个+1编程的核糖体框架转移事件.
  • 框架转移产生了一个新的C端域,形成一个α螺旋,缓解PLEKHM2.2的自身抑制.
  • 框架转移的PLEKHM2与基因素-1结合,用于传输到细胞尖端,独立于ARL8.
  • 需要恢复正规和框架转移的PLEKHM2蛋白质,以挽救淘汰心肌细胞的收缩功能.

结论:

  • 编程的核糖体框架转移对于PLEKHM2功能和正常的心脏活动至关重要.
  • 这一发现扩大了核编码基因中已知的框架转移机制的范围.
  • PLEKHM2的取决于移的调节会影响其局部化和细胞功能,特别是在心肌细胞中.