title: Acetamido sugar biosynthesis in the Euryarchaea.
abstract: Archaea and eukaryotes share a dolichol phosphate-dependent system for protein N-glycosylation. In both domains, the acetamido sugar N-acetylglucosamine (GlcNAc) forms part of the core oligosaccharide. However, the archaeal Methanococcales produce GlcNAc using the bacterial biosynthetic pathway. Key enzymes in this pathway belong to large families of proteins with diverse functions; therefore, the archaeal enzymes could not be identified solely using comparative sequence analysis. Genes encoding acetamido sugar-biosynthetic proteins were identified in Methanococcus maripaludis using phylogenetic and gene cluster analyses. Proteins expressed in Escherichia coli were purified and assayed for the predicted activities. The MMP1680 protein encodes a universally conserved glucosamine-6-phosphate synthase. The MMP1077 phosphomutase converted alpha-D-glucosamine-6-phosphate to alpha-D-glucosamine-1-phosphate, although this protein is more closely related to archaeal pentose and glucose phosphomutases than to bacterial glucosamine phosphomutases. The thermostable MJ1101 protein catalyzed both the acetylation of glucosamine-1-phosphate and the uridylyltransferase reaction with UTP to produce UDP-GlcNAc. The MMP0705 protein catalyzed the C-2 epimerization of UDP-GlcNAc, and the MMP0706 protein used NAD(+) to oxidize UDP-N-acetylmannosamine, forming UDP-N-acetylmannosaminuronate (ManNAcA). These two proteins are similar to enzymes used for proteobacterial lipopolysaccharide biosynthesis and gram-positive bacterial capsule production, suggesting a common evolutionary origin and a widespread distribution of ManNAcA. UDP-GlcNAc and UDP-ManNAcA biosynthesis evolved early in the euryarchaeal lineage, because most of their genomes contain orthologs of the five genes characterized here. These UDP-acetamido sugars are predicted to be precursors for flagellin and S-layer protein modifications and for the biosynthesis of methanogenic coenzyme B.
authors.affiliation: Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, Texas 78712, USA.
authors.forename: Seema C
authors.initials: SC
authors.lastname: Namboori
authors.forename: David E
authors.initials: DE
authors.lastname: Graham
mesh_terms: D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000096:biosynthesis* / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D053898:Biosynthetic Pathways; D002237:Carbohydrate Dehydrogenases / Q000378:metabolism; D002238:Carbohydrate Epimerases / Q000378:metabolism; D003001:Cloning, Molecular; D019641:DNA, Archaeal / Q000737:chemistry / Q000235:genetics; D004798:Enzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005944:Glucosamine / Q000031:analogs & derivatives / Q000378:metabolism; D019298:Glucose-6-Phosphate / Q000031:analogs & derivatives / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing) / Q000235:genetics / Q000378:metabolism; D017017:Methanococcus / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism; D017875:Phosphotransferases (Phosphomutases) / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism; D014539:Uridine Diphosphate Sugars / Q000378:metabolism; D014544:Uridine Triphosphate / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D004798:Enzymes; D005958:Glucosephosphates; D014539:Uridine Diphosphate Sugars; D009243:NAD; C038515:glucosamine 1-phosphate; C020949:uridine diphosphate N-acetyl-D-mannosaminuronic acid; C001293:glucosamine 6-phosphate; D014537:Uridine Diphosphate N-Acetylglucosamine; D019298:Glucose-6-Phosphate; C026146:N-acetylglucosamine-1-phosphate; D002237:Carbohydrate Dehydrogenases; C023228:UDP-N-acetylmannosamine dehydrogenase; D005945:Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing); D002238:Carbohydrate Epimerases; C100033:UDP acetylglucosamine-2-epimerase; D017875:Phosphotransferases (Phosphomutases); D005944:Glucosamine; D014544:Uridine Triphosphate; D000117:Acetylglucosamine