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Roles of key amino acid residues of the virulence factor listeriolysin O inListeria monocytogenes infection
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Yuchao FENG1, Jiali XU1, Binjie ZHU1, Zebin WU1, Wenjing WANG1, An DING1, Zimeng CAI1, Mianmian CHEN1, Jing SUN1, Lingli JIANG2, Houhui SONG1, Jing XIA1, *, Changyong CHENG1, *
Acta Microbiologica Sinica | 2024, 64(4) : 1219 - 1232
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Acta Microbiologica Sinica | 2024, 64(4): 1219-1232
• Research Articles •
Roles of key amino acid residues of the virulence factor listeriolysin O inListeria monocytogenes infection
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Yuchao FENG1, Jiali XU1, Binjie ZHU1, Zebin WU1, Wenjing WANG1, An DING1, Zimeng CAI1, Mianmian CHEN1, Jing SUN1, Lingli JIANG2, Houhui SONG1, Jing XIA1, *, Changyong CHENG1, *
Affiliations
  • 1 Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics & Advanced Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
  • 2 Ningbo College of Health Sciences, Ningbo 315100, Zhejiang, China
Published: 2024-04-04 doi: 10.13343/j.cnki.wsxb.20230679
Outline
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[Objective] To investigate the impacts of the amino acid residues at position 253 (glutamine, Q) and 254 (isoleucine, I) in the β8 sheet of the D3 domain of listeriolysin O (LLO) on the biological functions ofListeria monocytogenes. [Methods] We constructed the mutant proteins LLOQ253A and LLOI254A and the mutant strainshlyQ253A andhlyI254A by homologous recombination. After the expression and purification, the mutant proteins examined for the hemolytic activity. Furthermore, the growth, adhesion, invasion, intracellular migration, and proliferation were compared between the mutant strainshlyQ253A andhlyI254A. [Results] After the mutation of the corresponding sites, LLO proteins could be expressed normally. However, the mutant proteins and strains lost hemolytic activity at pH 6.5, and the hemolytic activities of LLOI254A andhlyI254A were restored at pH 5.5. The mutant strains showed no significant differences in extracellular growth, adhesion, and intracellular proliferation compared with the wild-type strain. However, the invasion and intercellular migration of the mutant strains were significantly lower than that of the wild-type strain. [Conclusion] The mutations of Q253A and I254A in LLO cause the loss of hemolytic activity at pH 6.5 and a reduction in the bacterial infection, the specific mechanisms of which remain to be explored. This study establishes a foundation for deeply understanding the impact of LLO structure on the biological function ofL.monocytogenes and holds significance for the construction of point-mutated strains ofL.monocytogenes.

Listeria monocytogenes  /  listeriolysin O  /  infection  /  point-mutated strains
Yuchao FENG, Jiali XU, Binjie ZHU, Zebin WU, Wenjing WANG, An DING, Zimeng CAI, Mianmian CHEN, Jing SUN, Lingli JIANG, Houhui SONG, Jing XIA, Changyong CHENG. Roles of key amino acid residues of the virulence factor listeriolysin O inListeria monocytogenes infection[J]. Acta Microbiologica Sinica, 2024 , 64 (4) : 1219 -1232 . DOI: 10.13343/j.cnki.wsxb.20230679
Funding
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  • National Natural Science Foundation of China(32172849)
  • National Natural Science Foundation of China(32302961)
  • National Science Foundation of Zhejiang Province(LQ22C180001)
  • National Science Foundation of Zhejiang Province(LY23C180002)
  • Ningbo Science and Technology Bureau(2022S006)
  • Zhejiang A&F University Talents Starting Program(2023LFR013)
Appendix
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Year 2024 volume 64 Issue 4
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144
67
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Article Info
doi: 10.13343/j.cnki.wsxb.20230679
  • Receive Date：2023-11-04
  • Online Date：2026-03-19
  • Published：2024-04-04
Article Data
Affiliations
History
  • Received：2023-11-04
  • Accepted：2024-01-05
Funding
National Natural Science Foundation of China(32172849)
National Natural Science Foundation of China(32302961)
National Science Foundation of Zhejiang Province(LQ22C180001)
National Science Foundation of Zhejiang Province(LY23C180002)
Ningbo Science and Technology Bureau(2022S006)
Zhejiang A&F University Talents Starting Program(2023LFR013)
Affiliations
    1 Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics & Advanced Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, College of Animal Science and Technology & College of Veterinary Medicine, Zhejiang A&F University, Hangzhou 311300, Zhejiang, China
    2 Ningbo College of Health Sciences, Ningbo 315100, Zhejiang, China

Corresponding:

*E-mail: XIA Jing,;
E-mail: CHENG Changyong,
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表12种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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