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Identification of Genes Involved in the Glycosylation of Modified Viosamine of Flagellins in Pseudomonas syringae by Mass Spectrometry.

Yamamoto M, Ohnishi-Kameyama M, Nguyen CL, Taguchi F, Chiku K, Ishii T, Ono H, Yoshida M, Ichinose Y - Genes (Basel) (2011)

Bottom Line: Previously we revealed that flagellin proteins in Pseudomonas syringae pv. tabaci 6605 (Pta 6605) were glycosylated with a trisaccharide, modified viosamine (mVio)-rhamnose-rhamnose and that glycosylation was required for virulence.We further identified some glycosylation-related genes, including vioA, vioB, vioT, fgt1, and fgt2.Furthermore, characterization of the mVio-related genes and MS analyses of flagellin glycans in other pathovars of P. syringae revealed that mVio-related genes were essential for mVio biosynthesis in flagellin glycans, and that P. syringae pv. syringae B728a, which does not possess a viosamine island, has a different structure of glycan in its flagellin protein.

View Article: PubMed Central - PubMed

Affiliation: National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. ymasanob@affrc.go.jp.

ABSTRACT
Previously we revealed that flagellin proteins in Pseudomonas syringae pv. tabaci 6605 (Pta 6605) were glycosylated with a trisaccharide, modified viosamine (mVio)-rhamnose-rhamnose and that glycosylation was required for virulence. We further identified some glycosylation-related genes, including vioA, vioB, vioT, fgt1, and fgt2. In this study, we newly identified vioR and vioM in a so-called viosamine island as biosynthetic genes for glycosylation of mVio in Pta 6605 by the mass spectrometry (MS) of flagellin glycan in the respective mutants. Furthermore, characterization of the mVio-related genes and MS analyses of flagellin glycans in other pathovars of P. syringae revealed that mVio-related genes were essential for mVio biosynthesis in flagellin glycans, and that P. syringae pv. syringae B728a, which does not possess a viosamine island, has a different structure of glycan in its flagellin protein.

No MeSH data available.


Related in: MedlinePlus

The metabolic pathway to produce modified viosamine and the structure of glycan attached to S201 in the flagellin (inset). Only the modified viosamine (in red) and the last precursor (in orange) were transferred to the non-reducing end of the rhamnosyl glycan by vioT.
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f3-genes-02-00788: The metabolic pathway to produce modified viosamine and the structure of glycan attached to S201 in the flagellin (inset). Only the modified viosamine (in red) and the last precursor (in orange) were transferred to the non-reducing end of the rhamnosyl glycan by vioT.

Mentions: The putative biosynthetic pathway of dTDP-mVio is illustrated in Figure 3. In a previous study we clarified that mVio was not transferred to the rhamnosyl glycan of flagellin in the mutants lacking vioA, vioB, and vioT [11]. Additionally, we revealed that the flagellin glycans in the vioR mutant were also composed of only rhamnose. On the other hand, the vioM mutant had demethylated mVio at the non-reducing end of flagellin glycans. These results indicated that the mVio-transfer enzyme, VioT, strictly recognized the modification of the amino group at position 4 of viosamine, and dTDP-N-acetoacetylviosamine and its precursors were not suitable for VioT as substrates. As a result, VioT recognizes dTDP-N-(3-hydroxy-1-oxobutyl)-2-O-methylviosamine and dTDP-N-(3-hydroxy-1-oxobutyl)-viosamine as the substrates and transfers N-(3-hydroxy-1-oxobutyl)-2-O-methylviosamine and N-(3-hydroxy-1-oxobutyl)-viosamine to the non-reducing terminus of rhamnosyl glycans.


Identification of Genes Involved in the Glycosylation of Modified Viosamine of Flagellins in Pseudomonas syringae by Mass Spectrometry.

Yamamoto M, Ohnishi-Kameyama M, Nguyen CL, Taguchi F, Chiku K, Ishii T, Ono H, Yoshida M, Ichinose Y - Genes (Basel) (2011)

The metabolic pathway to produce modified viosamine and the structure of glycan attached to S201 in the flagellin (inset). Only the modified viosamine (in red) and the last precursor (in orange) were transferred to the non-reducing end of the rhamnosyl glycan by vioT.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3927599&req=5

f3-genes-02-00788: The metabolic pathway to produce modified viosamine and the structure of glycan attached to S201 in the flagellin (inset). Only the modified viosamine (in red) and the last precursor (in orange) were transferred to the non-reducing end of the rhamnosyl glycan by vioT.
Mentions: The putative biosynthetic pathway of dTDP-mVio is illustrated in Figure 3. In a previous study we clarified that mVio was not transferred to the rhamnosyl glycan of flagellin in the mutants lacking vioA, vioB, and vioT [11]. Additionally, we revealed that the flagellin glycans in the vioR mutant were also composed of only rhamnose. On the other hand, the vioM mutant had demethylated mVio at the non-reducing end of flagellin glycans. These results indicated that the mVio-transfer enzyme, VioT, strictly recognized the modification of the amino group at position 4 of viosamine, and dTDP-N-acetoacetylviosamine and its precursors were not suitable for VioT as substrates. As a result, VioT recognizes dTDP-N-(3-hydroxy-1-oxobutyl)-2-O-methylviosamine and dTDP-N-(3-hydroxy-1-oxobutyl)-viosamine as the substrates and transfers N-(3-hydroxy-1-oxobutyl)-2-O-methylviosamine and N-(3-hydroxy-1-oxobutyl)-viosamine to the non-reducing terminus of rhamnosyl glycans.

Bottom Line: Previously we revealed that flagellin proteins in Pseudomonas syringae pv. tabaci 6605 (Pta 6605) were glycosylated with a trisaccharide, modified viosamine (mVio)-rhamnose-rhamnose and that glycosylation was required for virulence.We further identified some glycosylation-related genes, including vioA, vioB, vioT, fgt1, and fgt2.Furthermore, characterization of the mVio-related genes and MS analyses of flagellin glycans in other pathovars of P. syringae revealed that mVio-related genes were essential for mVio biosynthesis in flagellin glycans, and that P. syringae pv. syringae B728a, which does not possess a viosamine island, has a different structure of glycan in its flagellin protein.

View Article: PubMed Central - PubMed

Affiliation: National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. ymasanob@affrc.go.jp.

ABSTRACT
Previously we revealed that flagellin proteins in Pseudomonas syringae pv. tabaci 6605 (Pta 6605) were glycosylated with a trisaccharide, modified viosamine (mVio)-rhamnose-rhamnose and that glycosylation was required for virulence. We further identified some glycosylation-related genes, including vioA, vioB, vioT, fgt1, and fgt2. In this study, we newly identified vioR and vioM in a so-called viosamine island as biosynthetic genes for glycosylation of mVio in Pta 6605 by the mass spectrometry (MS) of flagellin glycan in the respective mutants. Furthermore, characterization of the mVio-related genes and MS analyses of flagellin glycans in other pathovars of P. syringae revealed that mVio-related genes were essential for mVio biosynthesis in flagellin glycans, and that P. syringae pv. syringae B728a, which does not possess a viosamine island, has a different structure of glycan in its flagellin protein.

No MeSH data available.


Related in: MedlinePlus