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Proc Natl Acad Sci U S A. 2010 Nov 23;107(47):20500-5. doi: 10.1073/pnas.0911253107. Epub 2010 Nov 5.

RNA polymerase mutants found through adaptive evolution reprogram Escherichia coli for optimal growth in minimal media.

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1
Department of Chemistry and Biochemistry, University of California at San Diego, La Jolla, CA 92093-0412, USA.

Abstract

Specific small deletions within the rpoC gene encoding the β'-subunit of RNA polymerase (RNAP) are found repeatedly after adaptation of Escherichia coli K-12 MG1655 to growth in minimal media. Here we present a multiscale analysis of these mutations. At the physiological level, the mutants grow 60% faster than the parent strain and convert the carbon source 15-35% more efficiently to biomass, but grow about 30% slower than the parent strain in rich medium. At the molecular level, the kinetic parameters of the mutated RNAP were found to be altered, resulting in a 4- to 30-fold decrease in open complex longevity at an rRNA promoter and a ∼10-fold decrease in transcriptional pausing, with consequent increase in transcript elongation rate. At a genome-scale, systems biology level, gene expression changes between the parent strain and adapted RNAP mutants reveal large-scale systematic transcriptional changes that influence specific cellular processes, including strong down-regulation of motility, acid resistance, fimbria, and curlin genes. RNAP genome-binding maps reveal redistribution of RNAP that may facilitate relief of a metabolic bottleneck to growth. These findings suggest that reprogramming the kinetic parameters of RNAP through specific mutations allows regulatory adaptation for optimal growth in new environments.

PMID:
21057108
PMCID:
PMC2996682
DOI:
10.1073/pnas.0911253107
[Indexed for MEDLINE]
Free PMC Article
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