Identification of genes involved in the biosynthesis and attachment of Methanococcus voltae N-linked glycans: insight into N-linked glycosylation pathways in Archaea.
N-linked glycosylation is recognized as an important post-translational modification across all three domains of life. However, the understanding of the genetic pathways for the assembly and attachment of N-linked glycans in eukaryotic and bacterial systems far outweighs the knowledge of comparable processes in Archaea. The recent characterization of a novel trisaccharide [beta-ManpNAcA6Thr-(1-4)-beta-GlcpNAc3NAcA-(1-3)-beta-GlcpNAc]N-linked to asparagine residues in Methanococcus voltae flagellin and S-layer proteins affords new opportunities to investigate N-linked glycosylation pathways in Archaea. In this contribution, the insertional inactivation of several candidate genes within the M. voltae genome and their resulting effects on flagellin and S-layer glycosylation are reported. Two of the candidate genes were shown to have effects on flagellin and S-layer protein molecular mass and N-linked glycan structure. Further examination revealed inactivation of either of these two genes also had effects on flagella assembly. These genes, designated agl (archaeal glycosylation) genes, include a glycosyl transferase (aglA) involved in the attachment of the terminal sugar to the glycan and an STT3 oligosaccharyl transferase homologue (aglB) involved in the transfer of the complete glycan to the flagellin and S-layer proteins. These findings document the first experimental evidence for genes involved in any glycosylation process within the domain Archaea.
Chaban
Bonnie
B
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, K7L 3N6, Canada.
Voisin
Sebastien
S
Kelly
John
J
Logan
Susan M
SM
Jarrell
Ken F
KF
D001105:Archaea / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid; D002874:Chromosome Mapping; D004591:Electrophoresis, Polyacrylamide Gel; D005408:Flagellin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D023061:Gene Order; D019766:Genes, Archaeal / Q000235:genetics; D006031:Glycosylation; D013058:Mass Spectrometry; D017017:Methanococcus / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D009844:Oligosaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000096:biosynthesis*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics
D019843:Archaeal Proteins; D009844:Oligosaccharides; D011134:Polysaccharides; D011994:Recombinant Proteins; D005408:Flagellin