In situ single-molecule dynamics of the SOS-repressor LexA during antibiotic stress
- Date
- Oct 15, 2020
- Time
- 11:00 AM - 12:00 PM
- Speaker
- Michael Schlierf
- Affiliation
- Technische Universität Dresden, Center for Molecular and Cellular Bioengineering (CMCB) B CUBE, Germany
- Series
- MPI-CBG Thursday Seminar
- Language
- en
- Main Topic
- Biologie
- Host
- Stefan Diez
- Description
- In bacteria, the key mechanism facilitating survival, mutation and adaptation upon DNA damage is the SOS response. Through autoproteolytic digestion triggered by single-stranded DNA, the repressor LexA controls over 50 SOS genes. Efforts to inhibit this response and thereby combat antibiotic resistance rely on a broad understanding of its behavior in vivo, which is still limited. Here, we use single-molecule localization microscopy to directly visualize LexA in Escherichia coli. With single-particle tracking PALM, we observe the repressor under antibiotic stress of different intensity and identify four diffusive populations. Using a set of LexA mutants, we assign operator bound, target-searching, freely diffusing and cleaved repressors. Measuring their time-evolution and applying a kinetic model, we obtain in vivo cleavage and binding rates in dependence of antibiotic dosage. A strong SOS response is even observed at low antibiotic stress. We further develop a strategy to count molecules in fixed cells and found that LexA is much more abundant during the late SOS response than previously thought.
Last modified: Oct 16, 2020, 12:09:58 AM
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Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG Auditorium (big half))Pfotenhauerstraße10801307Dresden
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- +49 351 210-0
- Fax
- +49 351 210-2000
- MPI-CBG
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- http://www.mpi-cbg.de
Organizer
Max Planck Institute of Molecular Cell Biology and GeneticsPfotenhauerstraße10801307Dresden
- Phone
- +49 351 210-0
- Fax
- +49 351 210-2000
- MPI-CBG
- Homepage
- http://www.mpi-cbg.de
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