Abstract
Gene gain by horizontal gene transfer is a major pathway of genome innovation in bacteria. The current view posits that acquired genes initially need to be silenced and that a bacterial chromatin protein, H-NS, plays a role in this silencing. However, we lack direct observation of the early fate of a horizontally transferred gene to prove this theory. We combine sequencing, flow cytometry and sorting, followed by microscopy to monitor gene expression and its variability after large-scale random insertions of a reporter gene in a population of Escherichia coli bacteria. We find that inserted promoters have a wide range of gene-expression variability related to their location. We find that high-expression clones carry insertions that are not correlated with H-NS binding. Conversely, binding of H-NS correlates with silencing. Finally, while most promoters show a common level of extrinsic noise, some insertions show higher noise levels. Analysis of these high-noise clones supports a scenario of switching due to transcriptional interference from divergent ribosomal promoters. Altogether, our findings point to evolutionary pathways where newly-acquired genes are not necessarily silenced, but may immediately explore a wide range of expression levels to probe the optimal ones.
Highlights
The high fraction of mobile genes in bacterial genomes is a source of a great diversity of phenotypes
The promoter chosen to control GFP expression in the randomly inserted cassette is the rrnBP1 promoter of the rrnB ribosomal operon. We chose this well-characterized highly expressed promoter because it is regulated by changes in DNA supercoiling and by the abundant nucleoid proteins Fis and H-NS [30]
We found an enrichment on H-NS binding sites in the surroundings (10 kb regions) of the insertions compared to random sites (Supplementary Figure S2)
Summary
The high fraction of mobile genes in bacterial genomes is a source of a great diversity of phenotypes.
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