Dissociated linkage of cytokine-inducing activity and cytotoxicity to different domains of listeriolysin O from Listeria monocytogenes.
Listeriolysin O (LLO), a cholesterol-binding cytolysin of Listeria monocytogenes, exhibits cytokine-inducing and cytolytic activities. Because the cytolytic activity was abolished by cholesterol treatment but the cytokine-inducing activity was not, these activities appeared to be linked to different domains of the LLO molecule. In this study, we constructed recombinant full-length LLO (rLLO529) and various truncated derivatives and examined their cytolytic, cholesterol-binding, and gamma interferon (IFN-gamma)-inducing activities. rLLO529 exhibited both IFN-gamma-inducing and cytolytic activities. Four truncated rLLOs possessing different C termini, which did not exert either cytolytic or cholesterol-binding activity, stimulated IFN-gamma production in normal spleen cells. However, a truncated rLLO corresponding to domain 4 (rLLO416-529) did not exhibit IFN-gamma-inducing activity, whereas it did bind to immobilized cholesterol. In addition, though the hemolysis induced by rLLO529 was inhibited by rLLO416-529, such inhibition was not detected upon rLLO529-induced IFN-gamma production. These data indicated that domain 4 was responsible for binding of LLO to membrane cholesterol followed by oligomerization and pore formation by the entire LLO molecule. In contrast, the other part of LLO, corresponding to domain 1-3, was essential for IFN-gamma-inducing activity. These findings implied a novel aspect of the function of LLO as a bacterial modulin.
Kohda
Chikara
C
Department of Microbiology, Kyoto University Graduate School of Medicine, Kyoto 606-8501,USA.
Kawamura
Ikuo
I
Baba
Hisashi
H
Nomura
Takamasa
T
Ito
Yutaka
Y
Kimoto
Terumi
T
Watanabe
Isao
I
Mitsuyama
Masao
M
D000818:Animals; D001427:Bacterial Toxins; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D003603:Cytotoxins / Q000235:genetics / Q000494:pharmacology*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000494:pharmacology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000494:pharmacology*; D007371:Interferon-gamma / Q000096:biosynthesis*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D051379:Mice; D008809:Mice, Inbred C3H; D010196:Pancreatic Elastase; D010446:Peptide Fragments / Q000494:pharmacology; D017434:Protein Structure, Tertiary; D012313:RNA; D018675:Toxicity Tests
D001427:Bacterial Toxins; D002352:Carrier Proteins; D003603:Cytotoxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D010446:Peptide Fragments; C039919:RNA, recombinant; D012313:RNA; D007371:Interferon-gamma; D002784:Cholesterol; D010196:Pancreatic Elastase; C017130:cholesterol-binding protein; C038705:hlyA protein, Listeria monocytogenes