Furthermore, there is evidence that transcripts can be cleaved within the A-site of paused or stalled ribosome (Hayes & Sauer, 2003) and such cleavage may lead to the triggering of trans-translation (Moore & Sauer, 2007). Thus, the role of trans-translation in reducing the effects of antimicrobial agents may relate more to overcoming the consequences of translational errors and truncated mRNA than to the stalled state caused by direct ribosome inhibition. As well as exposure to antimicrobial agents, there are several reports indicating that tmRNA levels can increase
under other conditions. For instance, increased levels of tmRNA correlated with G1–S transition in the cell cycle in Caulobacter crescentus
(Keiler & Shapiro, 2003; Hong et al., 2005) and in response to heat or chemical stress in Bacillus subtilis (Muto et al., 2000). In the former study (Keiler & Shapiro, 2003), the changing level of tmRNA Anti-diabetic Compound high throughput screening was believed to be critical to the timing of the click here cell cycle. In bacteria, mature tmRNA is one of the most abundant RNA species. tmRNA levels in M. smegmatis are equivalent to those reported for E. coli (Lee et al., 1978; Moore & Sauer, 2005). The abundance of tmRNA is a likely consequence of a high rate of trans-translation; for instance, approximately 0.4% of translation reactions in E. coli are terminated by trans-translation (Moore & Sauer, 2005). The abundance of tmRNA is also likely a consequence of its stability, which is believed to result from its binding to SmpB (Keiler et al., 2000; Moore & Sauer, 2005) and it is assumed that the majority of mature tmRNA and SmpB is in complex (Keiler, 2008). The half-life of mycobacterial tmRNA under conditions inhibiting RNA synthesis was similar to that reported for Caulobacter sp. swarmer cells and E. coli (Keiler & Shapiro, 2003). The stability of mycobacterial tmRNA was somewhat paradoxical in light of the high level of ssrA promoter Anacetrapib activity indicated by the results presented here. However, a previous study of the ssrA promoter of
C. crescentus also indicated that it was one of the most active promoters even under conditions where tmRNA was highly stable (Keiler & Shapiro, 2003). Irrespective of whether high-level ssrA promoter activity maintains tmRNA levels in the absence of ribosome inhibition, the evidence indicated that drug-associated increased levels of tmRNA were the result of increased promoter activity. This interpretation was supported not only by the promoter analysis but also by the finding that tmRNA loss was not affected by the drug exposure. The results presented here indicate that ribosome inhibitors, such as erythromycin, increase the synthesis of tmRNA in mycobacteria and thus provide an underlying mechanism for the increased levels of tmRNA following exposure to such agents.