A Defective Undecaprenyl Pyrophosphate Synthase Induces Growth and Morphological Defects That Are Suppressed by Mutations in the Isoprenoid Pathway of Escherichia coli

作者：MacCain William J; Kannan Suresh; Jameel Dannah Z; Troutman Jerry M; Young Kevin D

来源：Journal of Bacteriology, 2018, 200(18): e00255-18.

DOI：10.1128/JB.00255-18

• UppS
• isoprenoid
• undecaprenyl phosphate
• peptidoglycan
• morphology
• Und-P
• isoprenoids

The peptidoglycan exoskeleton shapes bacteria and protects them against osmotic forces, making its synthesis the target of many current antibiotics. Peptidoglycan precursors are attached to a lipid carrier and flipped from the cytoplasm into the periplasm to be incorporated into the cell wall. In Escherichia coil, this carrier is undecaprenyl phosphate (Und-P), which is synthesized as a diphosphate by the enzyme undecaprenyl pyrophosphate synthase (UppS). E. coil MG1655 exhibits wild-type morphology at all temperatures, but one of our laboratory strains (CS109) was highly aberrant when grown at 42 degrees C. This strain contained mutations affecting the Und-P synthetic pathway genes uppS, ispH, and idi. Normal morphology was restored by overexpressing uppS or by replacing the mutant (uppS31) with the wildtype allele. Importantly, moving uppS31 into MG1655 was lethal even at 30 degrees C, indicating that the altered enzyme was highly deleterious, but growth was restored by adding the CS109 versions of ispH and idi. Purified UppS(w)(31), was enzymatically defective at all temperatures, suggesting that it could not supply enough Und-P during rapid growth unless suppressor mutations were present. We conclude that cell wall synthesis is profoundly sensitive to changes in the pool of polyisoprenoids and that isoprenoid homeostasis exerts a particularly strong evolutionary pressure.
IMPORTANCE Bacterial morphology is determined primarily by the overall structure of the semirigid macromolecule peptidoglycan. Not only does peptidoglycan contribute to cell shape, but it also protects cells against lysis caused by excess osmotic pressure. Because it is critical for bacterial survival, it is no surprise that many antibiotics target peptidoglycan biosynthesis. However, important gaps remain in our understanding about how this process is affected by peptidoglycan precursor availability. Here, we report that a mutation altering the enzyme that synthesizes Und-P prevents cells from growing at high temperatures and that compensatory mutations in enzymes functioning upstream of uppS can reverse this phenotype. The results highlight the importance of Und-P metabolism for maintaining normal cell wall synthesis and shape.

• 出版日期2018-9