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Systematic dissection of σ⁷⁰ sequence diversity and function in bacteria.

ABSTRACT: Primary σ⁷⁰ factors are key conserved bacterial regulatory proteins that interact with regulatory DNA to control gene expression. It is, however, poorly understood whether σ⁷⁰ sequence diversity in different bacteria reflects functional differences. Here, we employ comparative and functional genomics to explore the sequence and function relationship of primary σ⁷⁰. Using multiplex automated genome engineering and deep sequencing (MAGE-seq), we generate a saturation mutagenesis library and high-resolution fitness map of E. coli σ⁷⁰ in domains 2-4. Mapping natural σ⁷⁰ sequence diversity to the E. coli σ⁷⁰ fitness landscape reveals significant predicted fitness deficits across σ⁷⁰ orthologs. Interestingly, these predicted deficits are larger than observed fitness changes for 15 σ⁷⁰ orthologs introduced into E. coli. Finally, we use a multiplexed transcriptional reporter assay and RNA sequencing (RNA-seq) to explore functional differences of several σ⁷⁰ orthologs. This work provides an in-depth analysis of σ⁷⁰ sequence and function to improve efforts to understand the evolution and engineering potential of this global regulator.

SUBMITTER: Park J

PROVIDER: S-EPMC8716302 | biostudies-literature |

REPOSITORIES: biostudies-literature

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