Real-time transposable element activity in individual live cells.
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.
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;
Neil H
0000-0001-9966-4998
NH
Kim
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;
Gloria
G
Lee
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;
Nicholas A
0000-0001-5940-2563
NA
Sherer
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;
K Michael
KM
Martini
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; tkuhlman@illinois.edu nigel@uiuc.edu.
Nigel
0000-0002-6322-0903
N
Goldenfeld
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; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801 tkuhlman@illinois.edu nigel@uiuc.edu.
Thomas E
0000-0002-4009-5601
TE
Kuhlman
D001426:Bacterial Proteins / Q000235:genetics; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics*; D005453:Fluorescence; D018628:Gene Dosage; D017930:Genes, Reporter; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids; D019895:Transposases / Q000235:genetics; D000097573:Red Fluorescent Protein
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D008164:Luminescent Proteins; D019895:Transposases; D000097573:Red Fluorescent Protein; C054562:yellow fluorescent protein, Bacteria
evolution; quantitative biology; transposable elements