title: Identification of multiple genes encoding membrane proteins with undecaprenyl pyrophosphate phosphatase (UppP) activity in Escherichia coli.
abstract: The bacA gene product of Escherichia coli was recently purified to near homogeneity and identified as an undecaprenyl pyrophosphate phosphatase activity (El Ghachi, M., Bouhss, A., Blanot, D., and Mengin-Lecreulx, D. (2004) J. Biol. Chem. 279, 30106-30113). The enzyme function is to synthesize the carrier lipid undecaprenyl phosphate that is essential for the biosynthesis of peptidoglycan and other cell wall components. The inactivation of the chromosomal bacA gene was not lethal but led to a significant, but not total, depletion of undecaprenyl pyrophosphate phosphatase activity in E. coli membranes, suggesting that other(s) protein(s) should exist and account for the residual activity and viability of the mutant strain. Here we report that inactivation of two additional genes, ybjG and pgpB, is required to abolish growth of the bacA mutant strain. Overexpression of either of these genes, or of a fourth identified one, yeiU, is shown to result in bacitracin resistance and increased levels of undecaprenyl pyrophosphate phosphatase activity, as previously observed for bacA. A thermosensitive conditional triple mutant delta bacA,delta ybjG,delta pgpB in which the expression of bacA is impaired at 42 degrees C was constructed. This strain was shown to accumulate soluble peptidoglycan nucleotide precursors and to lyse when grown at the restrictive temperature, due to the depletion of the pool of undecaprenyl phosphate and consequent arrest of cell wall synthesis. This work provides evidence that two different classes of proteins exhibit undecaprenyl pyrophosphate phosphatase activity in E. coli and probably other bacterial species; they are the BacA enzyme and several members from a superfamily of phosphatases that, different from BacA, share in common a characteristic phosphatase sequence motif.
authors.affiliation: Laboratoire des Enveloppes Bactériennes et Antibiotiques, Unite Mixte de Recherche 8619 CNRS, Université Paris-Sud, 91405 Orsay, France.
authors.forename: Meriem
authors.initials: M
authors.lastname: El Ghachi
authors.forename: Anne
authors.initials: A
authors.lastname: Derbise
authors.forename: Ahmed
authors.initials: A
authors.lastname: Bouhss
authors.forename: Dominique
authors.initials: D
authors.lastname: Mengin-Lecreulx
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001414:Bacitracin / Q000494:pharmacology; D001483:Base Sequence; D002384:Catalysis; D002462:Cell Membrane / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000502:physiology*; D017353:Gene Deletion; D005838:Genotype; D006358:Hot Temperature; D008055:Lipids / Q000737:chemistry; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D009841:Oligonucleotides / Q000737:chemistry; D010457:Peptidoglycan / Q000737:chemistry; D010641:Phenotype; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
chemical_list: D029968:Escherichia coli Proteins; D008055:Lipids; D009841:Oligonucleotides; D010457:Peptidoglycan; D011106:Polyisoprenyl Phosphates; D001414:Bacitracin; C088139:undecaprenyl pyrophosphate; C488524:BacA protein, E coli; D010744:Phosphoric Monoester Hydrolases