Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence

Kenta Imae, Yuki Saito, Hayato Kizaki, Hiroki Ryuno, Han Mao, Atsushi Miyashita, Yutaka Suzuki, Kazuhisa Sekimizu, Chikara Kaito

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We identified SA1684 as a Staphylococcus aureus virulence gene using a silkworm infection model. The SA1684 gene product carried the DUF402 domain, which is found in RNA-binding proteins, and had amino acid sequence similarity with a nucleoside diphosphatase, Streptomyces coelicolor SC4828 protein. The SA1684-deletion mutant exhibited drastically decreased virulence, in which the LD50 against silkworms was more than 10 times that of the parent strain. The SA1684-deletion mutant also exhibited decreased exotoxin production and colony-spreading ability. Purified SA1684 protein had Mn2+- or Co2+-dependent hydrolyzing activity against nucleoside diphosphates. Alanine substitutions of Tyr-88, Asp-106, and Asp-123/Glu-124, which are conserved between SA1684 and SC4828, diminished the nucleoside diphosphatase activity. Introduction of the wild-type SA1684 gene restored the hemolysin production of the SA1684-deletion mutant, whereas none of the alanine-substituted SA1684 mutant genes restored the hemolysin production. RNA sequence analysis revealed that SA1684 is required for the expression of the virulence regulatory genes agr, sarZ, and sarX, as well as metabolic genes involved in glycolysis and fermentation pathways. These findings suggest that the novel nucleoside diphosphataseSA1684links metabolic pathways and virulence gene expression and plays an important role in S. aureus virulence.

Original languageEnglish
Pages (from-to)18608-18619
Number of pages12
JournalJournal of Biological Chemistry
Volume291
Issue number36
DOIs
Publication statusPublished - Sep 2 2016
Externally publishedYes

Fingerprint

Virulence
Staphylococcus aureus
Genes
Hemolysin Proteins
Bombyx
Nucleosides
Alanine
Streptomyces coelicolor
RNA Sequence Analysis
Exotoxins
RNA-Binding Proteins
Diphosphates
Lethal Dose 50
Glycolysis
Regulator Genes
Metabolic Networks and Pathways
Fermentation
Amino Acid Sequence
Proteins
Gene expression

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence. / Imae, Kenta; Saito, Yuki; Kizaki, Hayato; Ryuno, Hiroki; Mao, Han; Miyashita, Atsushi; Suzuki, Yutaka; Sekimizu, Kazuhisa; Kaito, Chikara.

In: Journal of Biological Chemistry, Vol. 291, No. 36, 02.09.2016, p. 18608-18619.

Research output: Contribution to journalArticle

Imae, K, Saito, Y, Kizaki, H, Ryuno, H, Mao, H, Miyashita, A, Suzuki, Y, Sekimizu, K & Kaito, C 2016, 'Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence', Journal of Biological Chemistry, vol. 291, no. 36, pp. 18608-18619. https://doi.org/10.1074/jbc.M116.721845
Imae K, Saito Y, Kizaki H, Ryuno H, Mao H, Miyashita A et al. Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence. Journal of Biological Chemistry. 2016 Sep 2;291(36):18608-18619. https://doi.org/10.1074/jbc.M116.721845
Imae, Kenta ; Saito, Yuki ; Kizaki, Hayato ; Ryuno, Hiroki ; Mao, Han ; Miyashita, Atsushi ; Suzuki, Yutaka ; Sekimizu, Kazuhisa ; Kaito, Chikara. / Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 36. pp. 18608-18619.
@article{c8b800fca07745caa8b358a6466dc0df,
title = "Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence",
abstract = "We identified SA1684 as a Staphylococcus aureus virulence gene using a silkworm infection model. The SA1684 gene product carried the DUF402 domain, which is found in RNA-binding proteins, and had amino acid sequence similarity with a nucleoside diphosphatase, Streptomyces coelicolor SC4828 protein. The SA1684-deletion mutant exhibited drastically decreased virulence, in which the LD50 against silkworms was more than 10 times that of the parent strain. The SA1684-deletion mutant also exhibited decreased exotoxin production and colony-spreading ability. Purified SA1684 protein had Mn2+- or Co2+-dependent hydrolyzing activity against nucleoside diphosphates. Alanine substitutions of Tyr-88, Asp-106, and Asp-123/Glu-124, which are conserved between SA1684 and SC4828, diminished the nucleoside diphosphatase activity. Introduction of the wild-type SA1684 gene restored the hemolysin production of the SA1684-deletion mutant, whereas none of the alanine-substituted SA1684 mutant genes restored the hemolysin production. RNA sequence analysis revealed that SA1684 is required for the expression of the virulence regulatory genes agr, sarZ, and sarX, as well as metabolic genes involved in glycolysis and fermentation pathways. These findings suggest that the novel nucleoside diphosphataseSA1684links metabolic pathways and virulence gene expression and plays an important role in S. aureus virulence.",
author = "Kenta Imae and Yuki Saito and Hayato Kizaki and Hiroki Ryuno and Han Mao and Atsushi Miyashita and Yutaka Suzuki and Kazuhisa Sekimizu and Chikara Kaito",
year = "2016",
month = "9",
day = "2",
doi = "10.1074/jbc.M116.721845",
language = "English",
volume = "291",
pages = "18608--18619",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "36",

}

TY - JOUR

T1 - Novel nucleoside diphosphatase contributes to Staphylococcus aureus virulence

AU - Imae, Kenta

AU - Saito, Yuki

AU - Kizaki, Hayato

AU - Ryuno, Hiroki

AU - Mao, Han

AU - Miyashita, Atsushi

AU - Suzuki, Yutaka

AU - Sekimizu, Kazuhisa

AU - Kaito, Chikara

PY - 2016/9/2

Y1 - 2016/9/2

N2 - We identified SA1684 as a Staphylococcus aureus virulence gene using a silkworm infection model. The SA1684 gene product carried the DUF402 domain, which is found in RNA-binding proteins, and had amino acid sequence similarity with a nucleoside diphosphatase, Streptomyces coelicolor SC4828 protein. The SA1684-deletion mutant exhibited drastically decreased virulence, in which the LD50 against silkworms was more than 10 times that of the parent strain. The SA1684-deletion mutant also exhibited decreased exotoxin production and colony-spreading ability. Purified SA1684 protein had Mn2+- or Co2+-dependent hydrolyzing activity against nucleoside diphosphates. Alanine substitutions of Tyr-88, Asp-106, and Asp-123/Glu-124, which are conserved between SA1684 and SC4828, diminished the nucleoside diphosphatase activity. Introduction of the wild-type SA1684 gene restored the hemolysin production of the SA1684-deletion mutant, whereas none of the alanine-substituted SA1684 mutant genes restored the hemolysin production. RNA sequence analysis revealed that SA1684 is required for the expression of the virulence regulatory genes agr, sarZ, and sarX, as well as metabolic genes involved in glycolysis and fermentation pathways. These findings suggest that the novel nucleoside diphosphataseSA1684links metabolic pathways and virulence gene expression and plays an important role in S. aureus virulence.

AB - We identified SA1684 as a Staphylococcus aureus virulence gene using a silkworm infection model. The SA1684 gene product carried the DUF402 domain, which is found in RNA-binding proteins, and had amino acid sequence similarity with a nucleoside diphosphatase, Streptomyces coelicolor SC4828 protein. The SA1684-deletion mutant exhibited drastically decreased virulence, in which the LD50 against silkworms was more than 10 times that of the parent strain. The SA1684-deletion mutant also exhibited decreased exotoxin production and colony-spreading ability. Purified SA1684 protein had Mn2+- or Co2+-dependent hydrolyzing activity against nucleoside diphosphates. Alanine substitutions of Tyr-88, Asp-106, and Asp-123/Glu-124, which are conserved between SA1684 and SC4828, diminished the nucleoside diphosphatase activity. Introduction of the wild-type SA1684 gene restored the hemolysin production of the SA1684-deletion mutant, whereas none of the alanine-substituted SA1684 mutant genes restored the hemolysin production. RNA sequence analysis revealed that SA1684 is required for the expression of the virulence regulatory genes agr, sarZ, and sarX, as well as metabolic genes involved in glycolysis and fermentation pathways. These findings suggest that the novel nucleoside diphosphataseSA1684links metabolic pathways and virulence gene expression and plays an important role in S. aureus virulence.

UR - http://www.scopus.com/inward/record.url?scp=84984916665&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84984916665&partnerID=8YFLogxK

U2 - 10.1074/jbc.M116.721845

DO - 10.1074/jbc.M116.721845

M3 - Article

C2 - 27422825

AN - SCOPUS:84984916665

VL - 291

SP - 18608

EP - 18619

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 36

ER -