周期性拉伸增加培养的人成纤维细胞中的剪接噪声率。
Cyclic stretch increases splicing noise rate in cultured human fibroblasts.
作者信息
Uhl Michael, Mellert Kevin, Striegl Britta, Deibler Martin, Lamla Markus, Spatz Joachim P, Kemkemer Ralf, Kaufmann Dieter
机构信息
Institute of Human Genetics, University of Ulm, Albert Einstein Allee 11, D 89070 Ulm, Germany.
出版信息
BMC Res Notes. 2011 Oct 31;4:470. doi: 10.1186/1756-0500-4-470.
BACKGROUND
Mechanical forces are known to alter the expression of genes, but it has so far not been reported whether they may influence the fidelity of nucleus-based processes. One experimental approach permitting to address this question is the application of cyclic stretch to cultured human fibroblasts. As a marker for the precision of nucleus-based processes, the number of errors that occur during co-transcriptional splicing can then be measured. This so-called splicing noise is found at low frequency in pre-mRNA splicing.
FINDINGS
The amount of splicing noise was measured by RT-qPCR of seven exon skips from the test genes AATF, MAP3K11, NF1, PCGF2, POLR2A and RABAC1. In cells treated by altered uniaxial cyclic stretching for 18 h, a uniform and significant increase of splicing noise was found for all detectable exon skips.
CONCLUSION
Our data demonstrate that application of cyclic stretch to cultured fibroblasts correlates with a reduced transcriptional fidelity caused by increasing splicing noise.
背景
已知机械力会改变基因表达，但迄今为止，尚未有关于机械力是否会影响基于细胞核的过程的准确性的报道。一种能够解决这个问题的实验方法是对培养的人类成纤维细胞施加周期性拉伸。作为基于细胞核的过程准确性的标志物，可以测量共转录剪接过程中出现的错误数量。这种所谓的剪接噪声在mRNA前体剪接中以低频出现。
研究结果
通过对测试基因AATF、MAP3K11、NF1、PCGF2、POLR2A和RABAC1的七个外显子跳跃进行逆转录定量聚合酶链反应（RT-qPCR）来测量剪接噪声的量。在经过改变的单轴循环拉伸处理18小时的细胞中，所有可检测到的外显子跳跃的剪接噪声都出现了一致且显著的增加。
结论
我们的数据表明，对培养的成纤维细胞施加周期性拉伸与因剪接噪声增加导致的转录保真度降低相关。
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