Ex vivo proteomics of Campylobacter jejuni 81-176 reveal that FabG affects fatty acid composition to alter bacterial growth fitness in the chicken gut.
Campylobacter jejuni is one of the leading causes of foodborne gastrointestinal illness worldwide. Here we performed ex vivo proteomic analysis of C. jejuni 81-176 in chicken, a main reservoir for human infection. At 0, 1 and 4 weeks post-infection (p.i.) with the GFP-expressing 81-176 strain, inocula were recovered from chicken ceca by cell sorting using flow cytometry. iTRAQ-coupled 2D-LC-MS/MS analyses that detected 55 C. jejuni proteins, among which either 3 (FabG, HydB, CJJ81176_0876) or 7 (MscS, CetB, FlhF, PurH, PglJ, LpxC, Icd) proteins exhibited >1.4-fold-increased expression at 1 or 4 week(s) p.i. compared with those at 0 weeks p.i., respectively. Deletion of the fabG gene clearly decreased the proportion of bacterial unsaturated fatty acids (UFAs) and chicken colonization. The UFA proportion of the parental strain was not altered when grown at 42 °C. These findings suggest that FabG might play a pivotal role in UFA production, linked to bacterial adaptation in the poultry host. To our knowledge, this is the first example of ex vivo C. jejuni proteomics, in which fatty acid metabolism might affect bacterial adaptation to the chicken host.
Division of Biomedical Food Research, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan. Electronic address: hasakura@nihs.go.jp.
Hiroshi
H
Asakura
Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Inada 2-11, Obihiro, Hokkaido 080-8555, Japan.
Keiko
K
Kawamoto
Department of Animal Science, Tokyo University of Agriculture, Funako 1737, Atsugi, Kanagawa 243-0034, Japan.
Satoshi
S
Murakami
Division of Biomedical Food Research, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan.
Masato
M
Tachibana
Research Center for Animal Hygiene and Food Safety, Obihiro University of Agriculture and Veterinary Medicine, Inada 2-11, Obihiro, Hokkaido 080-8555, Japan.
Hisao
H
Kurazono
Department of Domestic Science, Kyoto Seibo College, Fukakusatayacho 1, Fushimi-ku, Kyoto, Kyoto 612-0878, Japan.
Sou-Ichi
S
Makino
Department of Fisheries, Tokai University, 3-20-1 Orido, Shimizu-ku, Shimizu-shi, Shizuoka 424-8610, Japan.
Shigeki
S
Yamamoto
Division of Biomedical Food Research, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan.
Shizunobu
S
Igimi
D000429:Alcohol Oxidoreductases / Q000032:analysis* / Q000235:genetics; D000818:Animals; D016123:Campylobacter jejuni / Q000737:chemistry* / Q000254:growth & development*; D002645:Chickens; D002853:Chromatography, Liquid; D003600:Cytosol / Q000737:chemistry; D005231:Fatty Acids, Unsaturated / Q000032:analysis*; D005434:Flow Cytometry; D041981:Gastrointestinal Tract / Q000382:microbiology*; D017353:Gene Deletion; D020543:Proteome / Q000032:analysis*; D053719:Tandem Mass Spectrometry; D013696:Temperature; D013997:Time Factors
D005231:Fatty Acids, Unsaturated; D020543:Proteome; D000429:Alcohol Oxidoreductases; C023744:acetoacetyl-CoA reductase
Campylobacter jejuni; Chicken colonization; Fatty acid biosynthesis; LC–MS