Experimental Evolution of Extreme Resistance to Ionizing Radiation in Escherichia coli after 50 Cycles of Selection

Author:

Bruckbauer Steven T.1,Trimarco Joseph D.12,Martin Joel3,Bushnell Brian3,Senn Katherine A.4,Schackwitz Wendy3,Lipzen Anna3,Blow Matthew3,Wood Elizabeth A.1,Culberson Wesley S.56,Pennacchio Christa3,Cox Michael M.1

Affiliation:

1. Department of Biochemistry, University of Wisconsin—Madison, Madison, Wisconsin, USA

2. Duke Molecular Genetics and Microbiology, Duke University School of Medicine, Durham, North Carolina, USA

3. DOE Joint Genome Institute, Walnut Creek, California, USA

4. Department of Biomolecular Chemistry, University of Wisconsin—Madison, Madison, Wisconsin, USA

5. Department of Medical Physics, School of Medicine and Public Health, University of Wisconsin—Madison, Madison, Wisconsin, USA

6. Department of Medical Physics, University of Wisconsin—Madison, Madison, Wisconsin, USA

Abstract

Some bacterial species exhibit astonishing resistance to ionizing radiation, with Deinococcus radiodurans being the archetype. As natural IR sources rarely exceed mGy levels, the capacity of Deinococcus to survive 5,000 Gy has been attributed to desiccation resistance. To understand the molecular basis of true extreme IR resistance, we are using experimental evolution to generate strains of Escherichia coli with IR resistance levels comparable to Deinococcus . Experimental evolution has previously generated moderate radioresistance for multiple bacterial species. However, these efforts could not take advantage of modern genomic sequencing technologies. In this report, we examine four replicate bacterial populations after 50 selection cycles. Genomic sequencing allows us to follow the genesis of mutations in populations throughout selection. Novel mutations affecting genes encoding DNA repair proteins and RNA polymerase enhance radioresistance. However, more contributors are apparent.

Funder

Department of Energy Joint Genome Institute

HHS | NIH | National Institute of General Medical Sciences

Publisher

American Society for Microbiology

Subject

Molecular Biology,Microbiology

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