Ireland, William T, Beeler, Suzannah M, Flores-Bautista, Emanuel, McCarty, Nicholas S, Röschinger, Tom, Belliveau, Nathan M, Sweredoski, Michael J, Moradian, Annie, Kinney, Justin B, Phillips, Rob (September 2020) Deciphering the regulatory genome of Escherichia coli, one hundred promoters at a time. eLife, 9. pp. 1-76. ISSN 2050-084X
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Abstract
Advances in DNA sequencing have revolutionized our ability to read genomes. However, even in the most well-studied of organisms, the bacterium Escherichia coli, for ≈65% of promoters we remain ignorant of their regulation. Until we crack this regulatory Rosetta Stone, efforts to read and write genomes will remain haphazard. We introduce a new method, Reg-Seq, that links massively parallel reporter assays with mass spectrometry to produce a base pair resolution dissection of more than a E. coli promoters in 12 growth conditions. We demonstrate that the method recapitulates known regulatory information. Then, we examine regulatory architectures for more than 80 promoters which previously had no known regulatory information. In many cases, we also identify which transcription factors mediate their regulation. This method clears a path for highly multiplexed investigations of the regulatory genome of model organisms, with the potential of moving to an array of microbes of ecological and medical relevance.
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