Temperature, among other environmental factors, influences the incidence and severity of many plant diseases. Likewise, numerous traits, including the expression of virulence factors, are regulated by temperature. Little is known about the underlying genetic determinants of thermoregulation in plant-pathogenic bacteria. Previously, we showed that the expression of both fliC (encoding flagellin) and syfA (encoding a nonribosomal polypeptide synthetase) was suppressed at high temperatures in Pseudomonas syringae. In this work, we used a high-throughput screen to identify mutations that conferred overexpression of syfA at elevated temperatures (28°C compared to 20°C). Two genes, Psyr_2474, encoding an acyl-coenzyme A (CoA) dehydrogenase, and Psyr_4843, encoding an ortholog of RppH, which in Escherichia coli mediates RNA turnover, contribute to thermoregulation of syfA. To assess the global role of rppH in thermoregulation in P. syringae, RNA sequencing was used to compare the transcriptomes of an rppH deletion mutant and the wild-type strain incubated at 20°C and 30°C. The disruption of rppH had a large effect on the temperature-dependent transcriptome of P. syringae, affecting the expression of 569 genes at either 20°C or 30°C but not at both temperatures. Intriguingly, RppH is involved in the thermoregulation of ribosome-associated proteins, as well as of RNase E, suggesting a prominent role of rppH on the proteome in addition to its effect on the transcriptome.
Temperature is an important environmental factor that influences many aspects of microbial physiology and profoundly affects an organism's ability to survive and reproduce (1). Since microorganisms must appropriately both perceive and respond to changes in temperature, they possess some form of thermoregulated gene expression. Certain temperature responses appear to be conserved across diverse bacterial species, such as the cold shock and heat shock responses (2, 3). The stimulatory signals and regulatory mechanisms of these shock responses are also largely conserved. However, in addition to the shock responses, which maintain cellular functions after large and rapid temperature shifts, most bacterial species also possess more specialized forms of thermoregulation whose role is not necessarily to return the cell to homeostasis but, rather, to reprogram cells for fitness in the altered environment. For example, animal pathogens often use host body temperature as a cue to express virulence factors (4-6). In this setting, members of many taxa suppress the production of flagellar genes since they encode immune-eliciting antigens (i.e., flagellin) (7). While the suppression of flagellin at host temperatures is common in such animal pathogens, the stages within the flagellar hierarchy and the mechanisms by which such regulation is achieved differ between organisms (cf. E. coli [8], Yersinia [9-11], and Listeria [12]). Common thermoregulated traits, such as motility, therefore do not necessarily have conserved mechanisms of regulation. This may be a consequence ...