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

Masanobu Yamamoto; Mayumi Ohnishi-Kameyama; Chi L Nguyen; Fumiko Taguchi; Kazuhiro Chiku; Tadashi Ishii; Hiroshi Ono; Mitsuru Yoshida; Yuki Ichinose

doi:10.3390/genes2040788

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

Genes (Basel). 2011 Oct 28;2(4):788-803. doi: 10.3390/genes2040788.

Authors

Masanobu Yamamoto¹, Mayumi Ohnishi-Kameyama², Chi L Nguyen³, Fumiko Taguchi⁴, Kazuhiro Chiku⁵, Tadashi Ishii⁶, Hiroshi Ono⁷, Mitsuru Yoshida⁸, Yuki Ichinose⁹

Affiliations

¹ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. ymasanob@affrc.go.jp.
² National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. kameyama@affrc.go.jp.
³ The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan. linhchiagi@yahoo.com.
⁴ The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan. ftaguchi@cc.okayama-u.ac.jp.
⁵ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. kchiku@affrc.go.jp.
⁶ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. tishii@affrc.go.jp.
⁷ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. ono@affrc.go.jp.
⁸ National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. mitsuru@affrc.go.jp.
⁹ The Graduate School of Natural Science and Technology, Okayama University, Tsushima-naka 1-1-1, Okayama 700-8530, Japan. yuki@cc.okayama-u.ac.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.