Indole Production Promotes Escherichia coli Mixed-Culture Growth with Pseudomonas aeruginosa by Inhibiting Quorum Signaling

Abstract

ABSTRACTIndole production byEscherichia coli, discovered in the early 20th century, has been used as a diagnostic marker for distinguishingE. colifrom other enteric bacteria. By using transcriptional profiling and competition studies with defined mutants, we show that cyclic AMP (cAMP)-regulated indole formation is a major factor that enablesE. coligrowth in mixed biofilm and planktonic populations withPseudomonas aeruginosa. Mutants deficient in cAMP production (cyaA) or the cAMP receptor gene (crp), as well as indole production (tnaA), were not competitive in coculture withP. aeruginosabut could be restored to wild-type competitiveness by supplementation with a physiologically relevant indole concentration.E. colisdiAmutants, which lacked the receptor for both indole andN-acyl-homoserine lactones (AHLs), showed no change in competitive fitness, suggesting that indole acted directly onP. aeruginosa. AnE. colitnaAmutant strain regained wild-type competiveness if grown withP. aeruginosaAHL synthase (rhlIandrhlI lasI) mutants. In contrast to the wild type,P. aeruginosaAHL synthase mutants were unable to degrade indole. Indole produced during mixed-culture growth inhibited pyocyanin production and other AHL-regulated virulence factors inP. aeruginosa. Mixed-culture growth withP. aeruginosastimulated indole formation inE. colicpdA, which is unable to regulate cAMP levels, suggesting the potential for mixed-culture gene activation via cAMP. These findings illustrate how indole, an early described feature ofE. colicentral metabolism, can play a significant role in mixed-culture survival by inhibiting quorum-regulated competition factors inP. aeruginosa.