Elongation factor P is required for EIIGlc translation in Corynebacterium glutamicum due to an essential polyproline motif

Translating ribosomes require elongation factor P (EF-P) to incorporate consecutive prolines (XPPX) into nascent peptide chains. The proteome of Corynebacterium glutamicum ATCC 13032 contains a total of 1,468 XPPX motifs, many of which are found in proteins involved in primary and secondary metabolism. We show here that synthesis of EIIGlc, the glucose-specific permease of the phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) encoded by ptsG, is strongly dependent on EF-P, as an efp deletion mutant grows poorly on glucose as sole carbon source. The amount of EIIGlc is strongly reduced in this mutant, which consequently results in a lower rate of glucose uptake. Strikingly, the XPPX motif is essential for the activity of EIIGlc, and substitution of the prolines leads to inactivation of the protein. Finally, translation of GntR2, a transcriptional activator of ptsG, is also dependent on EF-P. However, its reduced amount in the efp mutant can be compensated for by other regulators. These results reveal for the first time a translational bottleneck involving production of the major glucose transporter EIIGlc, which has implications for future strain engineering strategies.

Automated summary

Translation of the glucose-specific permease EIIGlc and the transcriptional activator GntR2 are both dependent on elongation factor P (EF-P). XPPX motifs play a crucial role in the activity of EIIGlc. Decreased levels of EIIGlc in an efp mutant can be compensated for by other regulators. These findings highlight a translational bottleneck in the production of EIIGlc with implications for future strain engineering strategies.