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

@article{citeulike:4197057, 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.}, author = {VanDyke, D. J. and Wu, J. and Ng, S. Y. and Kanbe, M. and Chaban, B. and Aizawa, S. and Jarrell, K. F.}, citeulike-article-id = {4197057}, citeulike-linkout-0 = {http://dx.doi.org/10.1128/JB.00474-08}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/18539748}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=18539748}, doi = {10.1128/JB.00474-08}, issn = {1098-5530}, journal = {Journal of bacteriology}, keywords = {archaea, flagella, glycosylation, pili}, month = {August}, number = {15}, pages = {5300--5307}, posted-at = {2009-03-20 08:50:54}, priority = {2}, title = {Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis.}, url = {http://dx.doi.org/10.1128/JB.00474-08}, volume = {190}, year = {2008} }

TY - JOUR ID - citeulike:4197057 L3 - citeulike-article-id:4197057 N2 - 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. IS - 15 JF - Journal of bacteriology SN - 1098-5530 EP - 5307 TI - Identification of a putative acetyltransferase gene, MMP0350, which affects proper assembly of both flagella and pili in the archaeon Methanococcus maripaludis. VL - 190 SP - 5300 KW - archaea KW - flagella KW - glycosylation KW - pili AU - VanDyke, DJ AU - Wu, J AU - Ng, SY AU - Kanbe, M AU - Chaban, B AU - Aizawa, S AU - Jarrell, KF PY - 2008/08// UR - http://dx.doi.org/10.1128/JB.00474-08 ER -