半乳糖基转移酶gttA基因缺失对4b型单增李斯特菌致病性的影响

doi:10.16431/j.cnki.1671-7236.2024.03.033

Abstract

Abstract: 【Objective】 This study was aimed to investigate the effect of deletion of galactosyltransferase gttA gene on the anchoring of surface proteins and pathogenicity of Listeria monocytogenes,so as to provide a reference to the pathogenic mechanism research of Listeria monocytogenes.【Method】 Homologous recombination methods were used to construct the gttA gene deletion strain, the growth and motility of the parental strain ScottA,the deletion strain ΔgttA and the complement strain CΔgttA were analyzed.The effects of gttA gene deletion on the anchoring of two surface virulence proteins InlB and ActA in Listeria monocytogenes were investigated by Western blotting.The adhesion and invasion test of colorectal adenocarcinoma cell line (Caco-2) and mouse infection test were used to evaluate the effect of gttA gene deletion on the adhesion and invasion ability and pathogenicity of Listeria monocytogenes.【Result】 The growth and motility of Listeria monocytogenes were not affected by the deletion of gttA gene.Western blotting results showed that after the deletion of gttA gene,the anchor quantities of virulence proteins InlB and ActA on the bacterial cell wall surface were extremely significantly decreased (P＜0.01).The results of cell adhesion and invasion assay showed that ΔgttA had an extremely significantly lower adhesion and invasion ability to Caco-2 cells than ScottA and CΔgttA (P＜0.01).The results of mouse infection test showed that the colonization ability of Listeria monocytogenes in liver and spleen of mice was extremely significantly or significantly weakened due to the deletion of gttA gene (P＜0.01 or P＜0.05).After the gttA gene was replenished,the level of adhesion and invasion in Caco-2 cells,and the colonization ability of the bacteria in liver and spleen were restored to the level of the parental strain ScottA.【Conclusion】 The deletion of gttA gene did not affect the growth and motility of Listeria monocytogenes,but affected the anchoring of the major virulence proteins on the surface of Listeria monocytogenes,which in turn weakened the ability of the bacterium to invade Caco-2 cell line and the pathogenicity of the bacterium in mice.

Key words: Listeria monocytogenes; galactosyltransferase; surface protein; pathogenicity

CLC Number:

S852.61⁺5

ZHENG Minghao, GUO Qian, LIU Fang, LIANG Xiongyan, LIU Jing, YANG Yuying, TIAN Guangming, FANG Chun. Effect of Galactosyltransferase gttA Gene Deletion on Pathogenicity of Listeria monocytogenes Serotype 4b[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(3): 1214-1222.

References

[1] SAUER J D, HERSKOVITS A A, SAUER J D, et al.Metabolism of the Gram-positive bacterial pathogen Listeria monocytogenes[J].Microbiology Spectrum, 2019, 7(4):10.
[2] DISSON O, MOURA A, LECUIT M.Making sense of the biodiversity and virulence of Listeria monocytogenes[J].Trends in Microbiology, 2021, 29(9):811-822.
[3] PIZARRO-CERDA J, COSSART-P. Listeria monocytogenes:Cell biology of invasion and intracellular growth[J].Microbiology Spectrum, 2018, 6(6):GPP3-0013-2018.
[4] OSEK J, WIECZOREK K.Listeria monocytogenes:How this pathogen uses its virulence mechanisms to infect the hosts[J].Pathogens, 2022, 11(12):1491.
[5] SUMRALL E T, KELLER A P, SHEN Y, et al.Structure and function of Listeria teichoic acids and their implications[J].Molecular Microbiology, 2020, 113(3):627-637.
[6] PILLICH H, PURI M, CHAKRABORTY T.ActA of Listeria monocytogenes and its manifold activities as an important Listeria virulence factor[J].Current Topics in Microbiology and Immunology, 2017, 399:113-132.
[7] YIN Y, YAO H, DOIJAD S, et al.A hybrid sub-lineage of Listeria monocytogenes comprising hypervirulent isolates[J].Nature Communitions, 2019, 10(1):4283.
[8] DUSSURGET O, PIZARRO-CERDA J, COSSART P.Molecular determinants of Listeria monocytogenes virulence[J].Annual Review of Microbiology, 2004, 58:587-610.
[9] TRAVIER L, LECUIT M.Listeria monocytogenes ActA:A new function for a 'classic' virulence factor[J].Current Opinion in Microbiology, 2014, 17:53-60.
[10] SPEARS P A, HAVELL E A, HAMRICK T S, et al.Listeria monocytogenes wall teichoic acid decoration in virulence and cell-to-cell spread[J].Molecular Microbiology, 2016, 101(5):714-730.
[11] SUMRALL E T, SCHEFER C R E, RISMONDO J, et al.Galactosylated wall teichoic acid, but not lipoteichoic acid, retains InlB on the surface of serovar 4b Listeria monocytogenes[J].Molecular Microbiology, 2020, 113(3):638-649.
[12] SUMRALL ET, SHEN Y, KELLER AP, et al.Phage resistance at the cost of virulence:Listeria monocytogenes serovar 4b requires galactosylated teichoic acids for InlB-mediated invasion[J].PLoS Pathogens, 2019, 15(10):e1008032.
[13] 吕洁婷,孙静,郑光辉,等.单核细胞增多性李斯特菌谷胱甘肽还原酶GR的生物学特性[J].微生物学报, 2021, 61(3):714-728. LYU J T, SUN J, ZHENG G H, et al.Characterization of the glutathione reductase from Listeria monocytogenes[J].Acta Microbiologica Sinica, 2021, 61(3):714-728.(in Chinese)
[14] 袁梅,吉俊芝,张钰,等.糖基转移酶GalU对单增李斯特菌致病性的影响[J].畜牧与兽医, 2022, 54(7):65-70. YUAN M, JI J Z, ZHANG Y, et al.Roles of glycosyltransferase GalU in pathogenicity of Listeria monocytogenes[J].Animal Husbandry&Veterinary Medicine, 2022, 54(7):65-70.(in Chinese)
[15] 郭骞,张钰,方小伟,等.糖基翻转酶gtcA基因缺失降低单增李斯特菌1/2a血清型菌株致病性的研究[J].中国畜牧兽医, 2022, 49(6):2298-2306. GUO Q, ZHANG Y, FANG X W, et al.Study on glycosyl-flippase gtcA gene deletion reduces the pathogenicity of Listeria monocytogenes serotype 1/2a[J].China Animal Husbandry&Veterinary Medicine, 2022, 49(6):2298-2306.(in Chinese)
[16] 汪国俊,黄金梅,季强,等.单增李斯特菌组氨酸激酶AgrC对毒力的影响[J].中国兽医科学, 2021, 51(10):1270-1278. WANG G J, HUANG J M, JI Q, et al.Role of histidine kinase AgrC in virulence of Listeria monocytogenes[J].Chinese Veterinary Science, 2021, 51(10):1270-1278.(in Chinese)
[17] 任静静,杨铭伟,陈云飞,等.食源性单增李斯特菌inlA/inlB/inlC基因缺失株的构建及其生物学特性分析[J].中国畜牧兽医, 2019, 46(1):45-53. REN J J, YANG M W, CHEN Y F, et al.Construction and characterization of inlA/inlB/inlC genes deletion mutant strain of food-borne Listeria monocytogenes[J]. China Animal Husbandry&Veterinary Medicine, 2019, 46(1):45-53.(in Chinese)
[18] PROMADEJ N, FIEDLER F, COSSART P, et al.Cell wall teichoic acid glycosylation in Listeria monocytogenes serotype 4b requires gtcA, a novel, serogroup-specific gene[J].Bacteriology, 1999, 181(2):418-425.
[19] RISMONDO J, HADDAD T F M, SHEN Y, et al.GtcA is required for LTA glycosylation in Listeria monocytogenes serovar 1/2a and Bacillus subtilis[J].Cell Surface, 2020, 19(6):100038.
[20] MEIRELES D, POMBINHO R, CARVALHO F, et al.Listeria monocytogenes wall teichoic acid glycosylation promotes surface anchoring of virulence factors, resistance to antimicrobial peptides, and decreased susceptibility to antibiotics[J].Pathogens, 2020, 9(4):290.
[21] SHEN Y, NAUJOKAS M, PARK M, et al.InIB-dependent internalization of Listeria is mediated by the Met receptor tyrosine kinase[J]. Cell, 2000, 103(3):501-510.
[22] HAMON M, BIERNE H, COSSART P.Listeria monocytogenes:A multifaceted model[J].Nature Review Microbiology, 2006, 4(6):423-434.
[23] CHENG C, JIANG L, MA T, et al.Carboxyl-terminal residues N478 and V479 required for the cytolytic activity of listeriolysin O play a critical role in Listeria monocytogenes pathogenicity[J].Frontiers in Immunology, 2017, 8:1439.
[24] FANG C, CAO T, CHENG C, et al.Activation of PrfA results in overexpression of virulence factors but does not rescue the pathogenicity of Listeria monocytogenes M7[J].Journal of Medical Microbiology, 2015, 64(8):818-827.
[25] GARCÍA-DEL P F, CALVO E, D'ORAZIO V, et al.Association of ActA to peptidoglycan revealed by cell wall proteomics of intracellular Listeria monocytogenes[J].The Journal of Biological Chemistry, 2011, 286(40):34675-34689.
[26] HA J, OH H, KIM S, et al.Effect of gene ActA on the invasion efficiency of Listeria monocytogenes, as observed in healthy and senescent intestinal epithelial cells[J].Journal of Microbiology and Biotechnology, 2018, 28(1):59-64.
[27] IRETON K, RIGANO L A, DOWD G C.Role of host GTPases in infection by Listeria monocytogenes[J].Cell Microbiology, 2014, 16(9):1311-1320.

Effect of Galactosyltransferase gttA Gene Deletion on Pathogenicity of Listeria monocytogenes Serotype 4b

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