Characterization of novel ADP-ribosyltransferase toxin from Salmonella which cause Salmonellosis in adult cattle.

• Project/Area Number: 25450434
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Veterinary medical science
• Research Institution: National Agriculture and Food Research Organization
• Principal Investigator: UCHIDA Ikuo 国立研究開発法人農業・食品産業技術総合研究機構, 動物衛生研究部門 細菌・寄生虫研究領域, 領域長 (70355204)
• Co-Investigator(Kenkyū-buntansha): TAMAMURA Yukino 国立研究開発法人・農業・食品産業技術総合研究機構, 動物衛生研究部門・細菌・寄生虫研究領域, 研究員 (90584384)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: サルモネラ症 / ADP-リボシル化毒素 / 病原性因子 / 百日咳毒素 / G蛋白質 / 複合体毒素 / サルモネラ / DT104 / ADP-リボシル化 / AB5型毒素 / A1B5毒素 / 成牛型サルモネラ症 / 生物活性 / 発現調節機構

Outline of Final Research Achievements

Salmonella Typhimurium DT104 produces pertussis-like toxin (ArtAB-DT104), which catalyzes ADP-ribosylation of G proteins. Here, we characterized ArtAB-DT104 and homologs in S. Worthington and S. bongori (ArtAB-SW and ArtAB-Sb, respectively). ArtABs comprise a A subunit and pentameric B subunits. The A subunits of ArtAB-SW and ArtAB-Sb, which exhibit ADP-ribosyltransferase activity, had high amino acid sequence identity with ArtAB-DT104, whereas the B subunits of ArtAB-Sb showed lower identity. Injection of ArtABs was fatal in mice; LD50 of ArtAB-DT104 and ArtAB-SW were lower than that of ArtAB-Sb, suggesting that the B subunit is important for ArtAB toxicity. ArtABs catalyzed ADP-ribosylation of Gi proteins in RAW264.7 cells and increased intracellular cyclic AMP levels. Unlike Pertussis toxin, ArtABs did not induce lymphocytosis, possibly due to differences in ADP-ribosylation of target G proteins. These results suggest the general importance of ArtABs for Salmonella virulence.

Research Products

• [Journal Article] A case study on Salmonella enterica serovar Typhimurium at a dairy farm associated with massive sparrow death (2016)
  • Author(s): Tamamura, Y., Uchida, I., Tanaka, K., Nakano, Y., Izumiya, H., Takahashi, T., Kikuchi, N.
  • Journal Title: Acta Veterinaria Scandinavica Volume: 58:23 Issue: 1 Pages: 1-4
  • DOI: 10.1186/s13028-016-0205-8
  • NAID: 120006666485
  • Peer Reviewed / Open Access
• [Journal Article] Rapid detection of Salmonella enterica serovar Typhimurium DT104 strains by the polymerase chain reaction. (2015)
  • Author(s): Yukawa, S., Tamura, Y., Tanaka, K., Uchida, I.
  • Journal Title: Acta Veterinaria Scandinavica Volume: 57:59 Issue: 1 Pages: 1-5
  • DOI: 10.1186/s13028-015-0143-x
  • NAID: 120006311927
  • Peer Reviewed / Open Access
• [Journal Article] Molecular typing of Salmonella enterica serovar Enteritidis isolates from domestic animals in Japan by multilocus variable-number tandem repeat analysis: evidence of clonal dissemination and replacement (2014)
  • Author(s): Kobayashi, A., Takahashi, S., Ono, M., Tanaka, K., Kishima, M., Akiba, M., Uchida, I.
  • Journal Title: Acta Veterinaria Scandinavica Volume: 56:31 Issue: 1
  • DOI: 10.1186/1751-0147-56-31
  • Peer Reviewed / Open Access
• [Journal Article] Complete nucleotide sequences of virulence-resistance plasmids carried by emerging multidrug-resistant Salmonella enterica serovar Typhimurium isolated from cattle in Hokkaido, Japan (2013)
  • Author(s): Tamamura, Y, 他
  • Journal Title: PLOS ONE Volume: 8 Issue: 10 Pages: e77644-e77644
  • DOI: 10.1371/journal.pone.0077644
  • Peer Reviewed
• [Presentation] Salmonella Typhimurium DT104が産生するArtABのマウスマクロファージ由来細胞RAW264.7に対する細胞内cAMP濃度上昇活性 (2015)
  • Author(s): 玉村雪乃、内田郁夫、田中聖
  • Organizer: 第158回日本獣医学術集会
  • Place of Presentation: 北里大学獣医学部 (青森県十和田市）
  • Year and Date: 2015-09-07
• [Presentation] Salmonellaが産生するADP-リボシル化毒素ArtABの性状 (2015)
  • Author(s): 玉村雪乃,内田郁夫
  • Organizer: 第88回日本細菌学会
  • Place of Presentation: 岐阜大学 （岐阜市）
  • Year and Date: 2015-03-26 – 2015-03-28
• [Presentation] Salmonellaにおける百日咳毒素様蛋白ArtABの産生およびその性状について (2014)
  • Author(s): 玉村雪乃,内田郁夫,田中聖
  • Organizer: 第157回日本獣医学会学術集会講演要旨集
  • Place of Presentation: 北海道大学 (札幌市）
  • Year and Date: 2014-09-09 – 2014-09-12
• [Presentation] 牛由来多剤耐性Salmonella Typhimurium PFGE VII型菌が保有する薬剤耐性病原性プラスミドの解析 (2013)
  • Author(s): 玉村 雪乃, 他
  • Organizer: 第80回日本細菌学会北海道支部学術総会
  • Place of Presentation: 東京農業大学生物産業学部