OLE RNA protects extremophilic bacteria from alcohol toxicity

Jason G. Wallace, Zhiyuan Zhou, Ronald R. Breaker

OLE (Ornate, Large, Extremophilic) RNAs represent a recently discovered non-coding RNA class found in extremophilic anaerobic bacteria, including certain human pathogens. OLE RNAs exhibit several unusual characteristics that indicate a potentially novel function, including exceptionally high expression and localization to cell membranes via interaction with a protein partner called OLE-associated protein (OAP). In the current study, new genetic and phenotypic characteristics of OLE RNA from Bacillus halodurans C-125 were established. OLE RNA is transcribed at high levels from its own promoter under normal growth conditions and the transcript is exceptionally stable compared to most other RNAs. Expression is increased by ∼ 7-fold when cells are exposed to near lethal concentrations of short-chain alcohols such as ethanol or methanol. Strains wherein the genes for OLE and/or OAP are deleted are more susceptible to growth inhibition by alcohol and also become more sensitive to cold. Normal growth characteristics can be restored by expressing the genes for OLE and OAP from plasmids or from elsewhere on the chromosome. Our findings confirm a functional link between OLE and OAP and reveal the importance of a large non-coding RNA in the response to alcohol-induced stress.