Monitoring F165 1 P-Like Fimbria Expression at the Single-Cell Level Reveals a Highly Heterogeneous Phenotype

Abstract

ABSTRACT F165 1 and the pyelonephritis-associated pili (Pap) are two members of the type P family of adhesive factors. They play a key role in establishing disease caused by extraintestinal pathogenic Escherichia coli (ExPEC) strains in animals and humans. Both F165 1 and Pap are under the control of an epigenetic and reversible switch that defines the number of fimbriated (ON) and afimbriated (OFF) cells within a clonal population. Using the Gfp reporter system, we monitored in vitro the level of fluorescence intensity corresponding to the F165 1 and Pap fimbrial synthesis. Monitoring individual Escherichia coli cells by flow cytometry and by real-time fluorescence microscopy, we identified cells associated with a low or high level of fluorescence intensity and a large amount of cells with partial levels of fluorescence, mostly present in the F165 1 system. This mixed population identified through fluorescence intensity could be attributed to the high switching rate previously observed in F165 1 -positive bacteria. The fimbrial heterogeneous phenotype for these ExPEC could represent increased fitness in unpredictable environments. Our study illustrates that within the large repertoire of fimbrial variants such as the well-characterized Pap, F165 1 is an exquisite example of regulatory expression that arms the bacterium with strategies for surviving in more than one particular environment.