首页 > 过刊浏览>2024年第43卷第6期 >289-296. DOI:10.13300/j.cnki.hnlkxb.2024.06.028
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N-乙酰转移酶对罗非鱼无乳链球菌毒力相关特性的影响
作者:
作者单位:

1.华中农业大学水产学院/湖北洪山实验室，武汉430070;2.岭南现代农业科学与技术广东省实验室，广州 510642

作者简介:

马天雨，E-mail：legenyu@163.com

通讯作者:

曾辉，E-mail：huizeng@mail.hzau.edu.cn

中图分类号:

S917.1

基金项目:

武汉市知识创新专项-基础研究项目（2023020201010103）；中央高校基本科研业务费专项（2662023PY021）；国家现代农业产业技术体系专项（CARS-46）；国家自然科学基金面上项目（32173015）；岭南现代农业实验室科研项目（NT2021008）


Effect of N-acetyltransferase on virulence properties of Streptococcus agalactiae
Author:
Affiliation:

1.College of Fisheries， Huazhong Agricultural University/Hubei Hongshan Laboratory， Wuhan 430070， China;2.Guangdong Laboratory for Lingnan Modern Agriculture， Guangzhou 510642， China

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    摘要:

    为阐明N-乙酰转移酶（NAT）在无乳链球菌致病过程中的作用，利用同源重组法构建野生株HN016的nat基因缺失株Δnat，并比较野生株和缺失株的生长、形态、黏附入侵细胞能力、抗吞噬能力、全血存活能力和致病力。结果显示，Δnat的溶血能力降低，并且nat缺失导致无乳链球菌对罗非鱼脑细胞的黏附和入侵能力下降，并增加了细菌对血液杀伤和吞噬细胞吞噬的敏感性。罗非鱼感染试验结果显示，nat基因缺失株Δnat（2×108 CFU/尾，致死率为36%）对罗非鱼的致死力显著低于野生株HN016（2×108 CFU/尾，致死率为85%）。罗非鱼感染12 h后，与野生对照株相比，Δnat突变株在血液、脾脏和脑组织中载菌量显著降低。研究表明，N-乙酰转移酶参与无乳链球菌抵抗宿主血液和吞噬细胞的清除，并协助细菌对罗非鱼组织的入侵和定植。

    Abstract:

    To explore the effect of N-acetyltransferase（NAT） on the pathogenicity of Streptococcus agalacae， the nat gene deletion strain Δnat was constructed by homologous recombination in the background of a wild strain HN016. The growth curve， morphology， cell adhesion， invasion， antiphagocytosis， whole blood survival and pathogenicity of wild strain and Δnat strain were compared. The results showed that the hemolytic capacity of Δnat decreased， and the loss of nat decreased the adhesion and invasion of S. agalactiae to tilapia brain cells and increased the bacterial susceptibility to blood killing and phagocytosis. The result of challenge experiment of tilapia showed that the lethality of Δnat （2×108 CFU/fish， the mortality rate was 36%） was significantly lower than that of wild strain HN016 （2×108 CFU/fish， the mortality rate was 85%）. And the bacterial loads of Δnat were much less than that of HN016 in blood， spleen and brain tissue at 12 h post infection. This study firstly reports that NAT plays an important role in S. agalactiae resistance to host blood and phagocyte clearance， and assists in the bacterial invasion and colonization of tilapia tissues.

    图1 Δnat缺失株验证Fig.1 Verification of the deletion strain Δnat
    图2 Δnat和野生株的生长曲线（A）、革兰氏染色（B）和溶血性（C）Fig.2 Growth curve （A）， Gram stain （B）， and hemolytic （C） of Δnat and wild type
    图3 Δnat和野生株在罗非鱼全血中生存能力Fig.3 Survival of Δnat and wild type in whole blood of tilapia
    图4 Δnat和野生株对罗非鱼脑细胞的黏附（A）和入侵能力（B）Fig.4 Adhesion（A） and invasion ability（B） of Δnat and wild type to tilapia brain cells
    图5 Δnat和野生株的抗吞噬能力Fig.5 Evaluation of anti-phagocytosis ability of Δnat and wild type
    图6 感染突变株和野生株后罗非鱼存活曲线（A）和症状（B）Fig.6 Tilapia survival curves （A） and symptoms （B） after infection with Δnat and wild type
    图7 罗非鱼组织载菌量Fig.7 Tissue bacterial load in tilapia
    表 1 引物信息Table 1 Primers information
    参考文献
    [1] CHEN M,LI L P,WANG R,et al.PCR detection and PFGE genotype analyses of streptococcal clinical isolates from tilapia in China[J].Veterinary microbiology,2012,159(3/4):526-530.
    [2] 陆雨星.氨基酸小分子变构调控ACT-GNAT型乙酰转移酶功能研究[D].上海:华东理工大学,2017.LU Y X.Study on the function of ACT-GNAT acetyltransferase regulated by amino acid small molecule allosteric regulation[D].Shanghai:East China University of Science and Technology,2017 (in Chinese with English abstract).
    [3] CASTA?O-CEREZO S,BERNAL V,POST H,et al.Protein acetylation affects acetate metabolism,motility and acid stress response in Escherichia coli[J/OL].Molecular systems biology,2014,10(11):762[2023-08-17]. https://doi.org/10.15252/msb.20145227.
    [4] SHIRMAST P,GHAFOORI S M,IRWIN R M,et al.Structural characterization of a GNAT family acetyltransferase from Elizabethkingia anophelis bound to acetyl-CoA reveals a new dimeric interface[J/OL].Scientific reports,2021,11(1):1274[2023-08-17].https://doi.org/10.1038/s41598-020-79649-5.
    [5] BERNDSEN C E,DENU J M.Catalysis and substrate selection by histone/protein lysine acetyltransferases[J].Current opinion in structural biology,2008,18(6):682-689.
    [6] CHEN W J,BISWAS T,PORTER V R,et al.Unusual regioversatility of acetyltransferase Eis,a cause of drug resistance in XDR-TB[J].PNAS,2011,108(24):9804-9808.
    [7] BURCKHARDT R M,ESCALANTE-SEMERENA J C.Insights into the function of the N-acetyltransferase SatA that detoxifies streptothricin in Bacillus subtilis and Bacillus anthracis[J/OL].Applied and environmental microbiology,2019,85(6):e03029-e03018[2023-08-17]. https://doi.org/10.1128/AEM.03029-18.
    [8] MISHRA A,ROY F,DOU Y T,et al.Role of acetyltransferase PG1842 in gingipain biogenesis in Porphyromonas gingivalis[J/OL].Journal of bacteriology,2018,200(24):e00385-e00318[2023-08-17]. https://doi.org/10.1128/jb.00385-18.
    [9] ALBERTí S,ASHBAUGH C D,WESSELS M R.Structure of the has operon promoter and regulation of hyaluronic acid capsule expression in group A Streptococcus[J].Molecular microbiology,1998,28(2):343-353.
    [10] LEWIS A L,HENSLER M E,VARKI A,et al.The group B streptococcal sialic acid O-acetyltransferase is encoded by neuD,a conserved component of bacterial sialic acid biosynthetic gene clusters[J].Journal of biological chemistry,2006,281(16):11186-11192.
    [11] DONS L,ERIKSSON E,JIN Y X,et al.Role of flagellin and the two-component CheA/CheY system of Listeria monocytogenes in host cell invasion and virulence[J].Infection and immunity,2004,72(6):3237-3244.
    [12] DONG Y H,MA K,CAO Q,et al.CRISPR-dependent endogenous gene regulation is required for virulence in piscine Streptococcus agalactiae[J].Emerging microbes & infections,2021,10(1):2113-2124.
    [13] HACKER J,HUGHES C.Genetics of Escherichia coli hemolysin[J] Current topics in microbiology and immunology,1985,118:139-162.
    [14] LEYING H,SUERBAUM S,KROLL H P,et al.The capsular polysaccharide is a major determinant of serum resistance in K-1-positive blood culture isolates of Escherichia coli[J].Infection and immunity,1990,58(1):222-227.
    [15] COLINO J,OUTSCHOORN I.The form variation of the capsular polysaccharide K1 is not a critical virulence factor of Escherichia coli in a neonatal mouse model of infection[J].Microbial pathogenesis,1999,27(4):187-196.
    [16] YAO R,BURR D H,GUERRY P.CheY-mediated modulation of Campylobacter jejuni virulence[J].Molecular microbiology,1997,23(5):1021-1031.
    [17] REN J,SANG Y,QIN R,et al.Metabolic intermediate acetyl phosphate modulates bacterial virulence via acetylation[J].Emerging microbes & infections,2019,8(1):55-69.
    [18] OUIDIR T,COSETTE P,JOUENNE T,et al.Proteomic profiling of lysine acetylation in Pseudomonas aeruginosa reveals the diversity of acetylated proteins[J].Proteomics,2015,15(13):2152-2157.
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马天雨,石富强,刘欣,张永安,曾辉. N-乙酰转移酶对罗非鱼无乳链球菌毒力相关特性的影响[J].华中农业大学学报,2024,43(6):289-296

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  • 收稿日期:2023-08-17
  • 在线发布日期: 2025-01-07
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