Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis

David J. VanDyke, John Wu, Sandy Y.M. Ng, Masaomi Kanbe, Bonnie Chaban, Shin Ichi Aizawa, Ken F. Jarrell

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Glycosylation is a posttranslational modification utilized in all three domains of life. Compared to eukaryotic and bacterial systems, knowledge of the archaeal processes involved in glycosylation is limited. Recently, Methanococcus voltae flagellin proteins were found to have an N-linked trisaccharide necessary for proper flagellum assembly. Current analysis by mass spectrometry of Methanococcus maripaludis flagellin proteins also indicated the attachment of an N-glycan containing acetylated sugars. To identify genes involved in sugar biosynthesis in M. maripaludis, a putative acetyltransferase was targeted for in-frame deletion. Deletion of this gene (MMP0350) resulted in a flagellin molecular mass shift to a size comparable to that expected for underglycosylated or completely nonglycoslyated flagellins, as determined by immunoblotting. Assembled flagellar filaments were not observed by electron microscopy. Interestingly, the deletion also resulted in defective pilus anchoring. Mutant cells with a deletion of MMP0350 had very few, if any, pili attached to the cell surface compared to a nonflagellated but piliated strain. However, pili were obtained from culture supernatants of this strain, indicating that the defect was not in pilus assembly but in stable attachment to the cell surface. Complementation of MMP0350 on a plasmid restored pilus attachment, but it was unable to restore flagellation, likely because the mutant ceased to make detectable flagellin. These findings represent the first report of a biosynthetic gene involved in flagellin glycosylation in archaea. Also, it is the first gene to be associated with pili, linking flagellum and pilus structure and assembly through posttranslational modifications.

Original languageEnglish
Pages (from-to)5300-5307
Number of pages8
JournalJournal of Bacteriology
Volume190
Issue number15
DOIs
Publication statusPublished - Aug 1 2008
Externally publishedYes

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Methanococcus
Flagellin
Acetyltransferases
Flagella
Archaea
Glycosylation
Genes
Post Translational Protein Processing
Trisaccharides
Gene Deletion
Immunoblotting
Polysaccharides
Mass Spectrometry
Electron Microscopy
Proteins
Plasmids

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis. / VanDyke, David J.; Wu, John; Ng, Sandy Y.M.; Kanbe, Masaomi; Chaban, Bonnie; Aizawa, Shin Ichi; Jarrell, Ken F.

In: Journal of Bacteriology, Vol. 190, No. 15, 01.08.2008, p. 5300-5307.

Research output: Contribution to journalArticle

VanDyke, David J. ; Wu, John ; Ng, Sandy Y.M. ; Kanbe, Masaomi ; Chaban, Bonnie ; Aizawa, Shin Ichi ; Jarrell, Ken F. / Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis. In: Journal of Bacteriology. 2008 ; Vol. 190, No. 15. pp. 5300-5307.
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