Journal article
Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis
Journal of Bacteriology, Vol.190(15), pp.5300-5307
2008
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. Copyright © 2008, American Society for Microbiology. All Rights Reserved.
Details
- Title
- Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis
- Authors
- D J VanDyke (Author) - Queen's University, CanadaJohn Wu (Author) - Queen's University, CanadaS Y M Ng (Author) - Queen's University, CanadaM Kanbe (Author) - Perfectural University of Hiroshima, JapanBonnie L Chaban (Author) - Queen's University, CanadaS I Aizawa (Author) - Perfectural University of Hiroshima, JapanK F Jarrell (Author) - Queen's University, Canada
- Publication details
- Journal of Bacteriology, Vol.190(15), pp.5300-5307
- Publisher
- American Society for Microbiology
- Date published
- 2008
- DOI
- 10.1128/JB.00474-08
- ISSN
- 0021-9193
- Organisation Unit
- University of the Sunshine Coast, Queensland; Thompson Institute; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 99450569602621
- Output Type
- Journal article
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