Kim Neil H, Lee Gloria, Sherer Nicholas A, Martini K Michael, Goldenfeld Nigel, Kuhlman Thomas E
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801; Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, IL 61801;
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801; Center for the Physics of Living Cells, University of Illinois at Urbana-Champaign, Urbana, IL 61801; Institute for Universal Biology NASA Astrobiology Institute, University of Illinois at Urbana-Champaign, Urbana, IL 61801; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801;
Proc Natl Acad Sci U S A. 2016 Jun 28;113(26):7278-83. doi: 10.1073/pnas.1601833113. Epub 2016 Jun 13.
The excision and reintegration of transposable elements (TEs) restructure their host genomes, generating cellular diversity involved in evolution, development, and the etiology of human diseases. Our current knowledge of TE behavior primarily results from bulk techniques that generate time and cell ensemble averages, but cannot capture cell-to-cell variation or local environmental and temporal variability. We have developed an experimental system based on the bacterial TE IS608 that uses fluorescent reporters to directly observe single TE excision events in individual cells in real time. We find that TE activity depends upon the TE's orientation in the genome and the amount of transposase protein in the cell. We also find that TE activity is highly variable throughout the lifetime of the cell. Upon entering stationary phase, TE activity increases in cells hereditarily predisposed to TE activity. These direct observations demonstrate that real-time live-cell imaging of evolution at the molecular and individual event level is a powerful tool for the exploration of genome plasticity in stressed cells.
转座元件(TEs)的切除和重新整合会重塑其宿主基因组,产生与进化、发育以及人类疾病病因相关的细胞多样性。我们目前对TE行为的了解主要源于大量技术,这些技术生成的是时间和细胞群体平均值,但无法捕捉细胞间的差异或局部环境及时间变异性。我们基于细菌转座元件IS608开发了一个实验系统,该系统使用荧光报告基因实时直接观察单个细胞中的单个TE切除事件。我们发现TE活性取决于TE在基因组中的方向以及细胞中转座酶蛋白的量。我们还发现,在细胞的整个生命周期中,TE活性高度可变。进入稳定期后,在遗传上易于发生TE活性的细胞中,TE活性会增加。这些直接观察结果表明,在分子和个体事件水平上对进化进行实时活细胞成像,是探索应激细胞中基因组可塑性的有力工具。