@article {Zarkan425538, author = {Ashraf Zarkan and Santiago Ca{\~n}o Mu{\~n}iz and Jinbo Zhu and Kareem Al Nahas and Jehangir Cama and Ulrich F. Keyser and David K. Summers}, title = {Indole Pulse Signalling Regulates the Cytoplasmic pH of E. coli in a Memory-Like Manner}, elocation-id = {425538}, year = {2018}, doi = {10.1101/425538}, publisher = {Cold Spring Harbor Laboratory}, abstract = {Bacterial cells are critically dependent upon pH regulation. Most proteins function over a limited pH range and the pH gradient across the bacterial cell membrane is central to energy production and transduction1. Here we demonstrate that indole plays a critical role in the regulation of the cytoplasmic pH of E. coli. Indole is an aromatic molecule with diverse signalling roles that in bacteria is produced from tryptophan by the enzyme tryptophanase (TnaA)2. Two modes of indole signalling have been described: persistent and pulse signalling. The latter is illustrated by the brief but intense elevation of intracellular indole during stationary phase entry3,4. We show that E. coli cells growing under conditions where no indole is produced maintain their cytoplasmic pH at 7.8 {\textpm} 0.2. In contrast, under conditions permitting indole production, pH is maintained at 7.2 {\textpm} 0.2. Experiments where indole was added experimentally to non-producing cultures showed that pH regulation results from pulse, rather than persistent, indole signalling. Furthermore, the application of an artificial pulse of either of two non-biological proton ionophores (DNP or CCCP) caused a similar effect, suggesting that the relevant property of indole in this context is its ability to conduct protons across the cytoplasmic membrane5. Additionally, we show that the effect of the indole pulse that occurs normally during stationary phase entry in rich medium remains as a {\textquotedblleft}memory{\textquotedblright} to maintain the correct cytoplasmic pH until entry into the next stationary phase. The indole-mediated reduction in cytoplasmic pH may explain why indole provides E. coli with a degree of protection against stresses, including some bactericidal antibiotics.}, URL = {https://www.biorxiv.org/content/early/2018/09/24/425538}, eprint = {https://www.biorxiv.org/content/early/2018/09/24/425538.full.pdf}, journal = {bioRxiv} }