PMID- 22949549
OWN - NLM
STAT- MEDLINE
DCOM- 20130314
LR  - 20211203
IS  - 1098-5522 (Electronic)
IS  - 0019-9567 (Print)
IS  - 0019-9567 (Linking)
VI  - 80
IP  - 11
DP  - 2012 Nov
TI  - Subtilase cytotoxin enhances Escherichia coli survival in macrophages by 
      suppression of nitric oxide production through the inhibition of NF-kappaB 
      activation.
PG  - 3939-51
LID - 10.1128/IAI.00581-12 [doi]
AB  - Subtilase cytotoxin (SubAB), which is produced by certain strains of 
      Shiga-toxigenic Escherichia coli (STEC), cleaves an endoplasmic reticulum (ER) 
      chaperone, BiP/Grp78, leading to induction of ER stress and caspase-dependent 
      apoptosis. SubAB alters the innate immune response. SubAB pretreatment of 
      macrophages inhibited lipopolysaccharide (LPS)-induced production of both 
      monocyte chemoattractant protein 1 (MCP-1) and tumor necrosis factor alpha (TNF-alpha). 
      We investigated here the mechanism by which SubAB inhibits nitric oxide (NO) 
      production by mouse macrophages. SubAB suppressed LPS-induced NO production 
      through inhibition of inducible NO synthase (iNOS) mRNA and protein expression. 
      Further, SubAB inhibited LPS-induced IkappaB-alpha phosphorylation and nuclear 
      localization of the nuclear factor-kappaB (NF-kappaB) p65/p50 heterodimer. Reporter gene 
      and chromatin immunoprecipitation (ChIP) assays revealed that SubAB reduced 
      LPS-induced NF-kappaB p65/p50 heterodimer binding to an NF-kappaB binding site on the 
      iNOS promoter. In contrast to the native toxin, a catalytically inactivated SubAB 
      mutant slightly enhanced LPS-induced iNOS expression and binding of NF-kappaB 
      subunits to the iNOS promoter. The SubAB effect on LPS-induced iNOS expression 
      was significantly reduced in macrophages from NF-kappaB1 (p50)-deficient mice, which 
      lacked a DNA-binding subunit of the p65/p50 heterodimer, suggesting that p50 was 
      involved in SubAB-mediated inhibition of iNOS expression. Treatment of 
      macrophages with an NOS inhibitor or expression of SubAB by E. coli increased E. 
      coli survival in macrophages, suggesting that NO generated by macrophages 
      resulted in efficient killing of the bacteria and SubAB contributed to E. coli 
      survival in macrophages. Thus, we hypothesize that SubAB might represent a novel 
      bacterial strategy to circumvent host defense during STEC infection.
FAU - Tsutsuki, Hiroyasu
AU  - Tsutsuki H
AD  - Departments of Molecular Infectiology, Chiba University, Chiba, Japan.
FAU - Yahiro, Kinnosuke
AU  - Yahiro K
FAU - Suzuki, Kotaro
AU  - Suzuki K
FAU - Suto, Akira
AU  - Suto A
FAU - Ogura, Kohei
AU  - Ogura K
FAU - Nagasawa, Sayaka
AU  - Nagasawa S
FAU - Ihara, Hideshi
AU  - Ihara H
FAU - Shimizu, Takeshi
AU  - Shimizu T
FAU - Nakajima, Hiroshi
AU  - Nakajima H
FAU - Moss, Joel
AU  - Moss J
FAU - Noda, Masatoshi
AU  - Noda M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120904
PL  - United States
TA  - Infect Immun
JT  - Infection and immunity
JID - 0246127
RN  - 0 (Endoplasmic Reticulum Chaperone BiP)
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Hspa5 protein, mouse)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (NF-kappa B)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase Type II)
RN  - EC 3.4.21.- (Subtilisins)
RN  - EC 3.4.21.- (subtilase cytotoxin, E coli)
SB  - IM
MH  - Animals
MH  - Cell Survival
MH  - Cells, Cultured
MH  - Endoplasmic Reticulum Chaperone BiP
MH  - Escherichia coli/*metabolism
MH  - Escherichia coli Proteins/*pharmacology
MH  - Gene Expression Regulation
MH  - Immunoblotting
MH  - Immunoprecipitation
MH  - Lipopolysaccharides/pharmacology
MH  - Macrophages/*drug effects/metabolism
MH  - Mice
MH  - NF-kappa B/*antagonists & inhibitors/metabolism
MH  - Nitric Oxide/biosynthesis
MH  - Nitric Oxide Synthase Type II/*genetics/metabolism
MH  - Real-Time Polymerase Chain Reaction
MH  - Signal Transduction
MH  - Subtilisins/*pharmacology
PMC - PMC3486033
EDAT- 2012/09/06 06:00
MHDA- 2013/03/15 06:00
PMCR- 2013/05/01
CRDT- 2012/09/06 06:00
PHST- 2012/09/06 06:00 [entrez]
PHST- 2012/09/06 06:00 [pubmed]
PHST- 2013/03/15 06:00 [medline]
PHST- 2013/05/01 00:00 [pmc-release]
AID - IAI.00581-12 [pii]
AID - 00581-12 [pii]
AID - 10.1128/IAI.00581-12 [doi]
PST - ppublish
SO  - Infect Immun. 2012 Nov;80(11):3939-51. doi: 10.1128/IAI.00581-12. Epub 2012 Sep 
      4.