The Escherichia coli homologue of yeast RER2, a key enzyme of dolichol synthesis, is essential for carrier lipid formation in bacterial cell wall synthesis.
We found in the Escherichia coli genome sequence a homologue of RER2, a Saccharomyces cerevisiae gene required for proper localization of an endoplasmic reticulum protein, and designated it rth (RER2 homologue). The disruption of this gene was lethal for E. coli. To reveal its biological function, we isolated temperature-sensitive mutants of the rth gene. The mutant cells became swollen and burst at the nonpermissive temperature, indicating that their cell wall integrity was defective. Further analysis showed that the mutant cells were deficient in the activity of cis-prenyltransferase, namely, undecaprenyl diphosphate synthase, a key enzyme of the carrier lipid formation of peptidoglycan synthesis. The cellular level of undecaprenyl phosphate was in fact markedly decreased in the mutants. These results are consistent with the fact that the Rer2 homologue of Micrococcus luteus shows undecaprenyl diphosphate synthase activity (N. Shimizu, T. Koyama, and K. Ogura, J. Biol. Chem. 273:19476-19481, 1998) and demonstrate that E. coli Rth is indeed responsible for the maintenance of cell wall rigidity. Our work on the yeast rer2 mutants shows that they are defective in the activity of cis-prenyltransferase, namely, dehydrodolichyl diphosphate synthase, a key enzyme of dolichol synthesis. Taking these data together, we conclude that the RER2 gene family encodes cis-prenyltransferase, which plays an essential role in cell wall biosynthesis in bacteria and in dolichol synthesis in eukaryotic cells and has been well conserved during evolution.
Kato
J
J
Department of Molecular Biology, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan.
Fujisaki
S
S
Nakajima
K
K
Nishimura
Y
Y
Sato
M
M
Nakano
A
A
D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D002473:Cell Wall / Q000378:metabolism*; D004286:Dolichols / Q000096:biosynthesis*; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D005057:Eukaryotic Cells; D005798:Genes, Bacterial; D020043:Genes, Essential; D009154:Mutation; D011106:Polyisoprenyl Phosphates / Q000096:biosynthesis*; D011387:Prokaryotic Cells
D004286:Dolichols; D011106:Polyisoprenyl Phosphates; C009621:undecaprenyl phosphate; D019883:Alkyl and Aryl Transferases; C023266:undecaprenyl pyrophosphate synthetase