Shewanella oneidensis内源性大质粒上ParE-ParD家族毒素-抗毒素系统(SO_A0088-A0087)的鉴定

doi:10.11978/2018022

热带海洋学报 ›› 2018, Vol. 37 ›› Issue (6): 104-111.doi: 10.11978/2018022CSTR: 32234.14.2018022

Shewanella oneidensis内源性大质粒上ParE-ParD家族毒素-抗毒素系统(SO_A0088-A0087)的鉴定

赵玄玉^1,²(), 施斐¹, 王晓雪^1,²()

1. 中国科学院热带海洋生物资源与生态重点实验室, 中国科学院南海海洋研究所, 广东广州 510301
2. 中国科学院大学, 北京 100049

收稿日期:2018-02-28 出版日期:2018-11-20 发布日期:2018-12-24
作者简介:
作者简介：赵玄玉(1993—), 女, 广东省珠海市人, 海洋生物学硕士, 从事毒素-抗毒素系统的功能研究。E-mail: zhaoxuanyu88@163.com
基金资助:
国家自然科学基金(31500025、31500093)

Characterization of a ParE-ParD toxin-antitoxin TA system (SO_A0087-A0088) on a megaplasmid of Shewanella oneidensis

Xuanyu ZHAO^1,²(), Fei SHI¹, Xiaoxue WANG^1,²()

1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2018-02-28 Online:2018-11-20 Published:2018-12-24
Supported by:
National Natural Science Foundation of China (31500025, 31500093)

摘要/Abstract

摘要：

毒素-抗毒素系统在原核生物中分布十分广泛, 在细菌的生命活动中扮演了重要的角色, 如维持水平基因转移元件的稳定性以及应对环境胁迫等。然而目前对于来自生态环境菌株中的毒素-抗毒素系统的研究仍较少。文章鉴定了自然水体环境来源的菌株Shewanella oneidensis MR-1 携带的内源性大质粒上的一对ParE-ParD家族Ⅱ型毒素-抗毒素系统SO_A0088-A0087。毒素SO_A0088在大肠杆菌以及原宿主S. oneidensis 内均具有明显的细胞毒性, 并导致细胞分裂存在缺陷。抗毒素SO_A0087能够完全拮抗毒素的毒性。同时, 凝胶电泳迁移率实验(electrophoretic mobility shift assay, EMSA)证实抗毒素SO_A0087能够结合自身的启动子。另外, 文章还通过易错突变的方法找到了毒素SO_A0088的3个毒性关键位点。

关键词: 毒素-抗毒素系统, ParE-ParD, Shewanella oneidensis, 质粒

Abstract:

Toxin-antitoxin (TA) systems are prevalent in prokaryotes. They play important roles in cell physiological activities including maintenance of mobile genetic elements and mediation of stress responses. However, studies of TA systems in the bioelectrochemically and ecologically important strain Shewanella oneidensis are limited. In this study, we characterized SO_A0087-A0088 on the megaplasmid of Shewanella oneidensis MR-1 as a typical type Ⅱ TA system that belongs to ParE-ParD family, and the overexpression of toxin A0088 leads to filamentous growth. We found that the antitoxin SO_A0087 could bind to promoter of SO_A0087-A0088 in vitro, and three amino acid residues of toxin SO_A0088 were identified to be critical for toxicity by random mutagenesis.

Key words: toxin-antitoxin system, ParE-ParD, Shewanella oneidensis, plasmid

中图分类号:

Q936

赵玄玉, 施斐, 王晓雪. Shewanella oneidensis内源性大质粒上ParE-ParD家族毒素-抗毒素系统(SO_A0088-A0087)的鉴定[J]. 热带海洋学报, 2018, 37(6): 104-111.

Xuanyu ZHAO, Fei SHI, Xiaoxue WANG. Characterization of a ParE-ParD toxin-antitoxin TA system (SO_A0087-A0088) on a megaplasmid of Shewanella oneidensis[J]. Journal of Tropical Oceanography, 2018, 37(6): 104-111.

图/表 5

图1

图2

图 3

图4

图5

参考文献 30

[1]	BARBOSA L C B, GARRIDO S S, GARCIA A, et al, 2010. Function inferences from a molecular structural model of bacterial ParE toxin[J]. Bioinformation, 4(10): 438-440.
[2]	CHAN W T, ESPINOSA M, YEO C C, 2016. Keeping the wolves at bay: Antitoxins of prokaryotic type II toxin-antitoxin systems[J]. Frontiers in Molecular Biosciences, 3: 9.
[3]	DALTON K M, CROSSON S, 2010. A conserved mode of protein recognition and binding in a ParD-ParE toxin-antitoxin complex[J]. Biochemistry, 49(10): 2205-2215.
[4]	DUPRILOT M, DECRE D, GENEL N, et al, 2017. Diversity and functionality of plasmid-borne VagCD toxin-antitoxin systems of Klebsiella pneumoniae[J]. Journal of Antimicrobial Chemotherapy, 72(5): 1320-1326.
[5]	FIEBIG A, CASTRO ROJAS C M, SIEGAL-GASKINS D, et al, 2010. Interaction specificity, toxicity and regulation of a paralogous set of ParE/RelE-family toxin-antitoxin systems[J]. Molecular Microbiology, 77(1): 236-251.
[6]	FISHMAN A, TAO YING, BENTLEY W E, et al, 2004. Protein engineering of toluene 4-monooxygenase of Pseudomonas mendocina KR1 for synthesizing 4-nitrocatechol from nitrobenzene[J]. Biotechnology and Bioengineering, 87(6): 779-790.
[7]	HALLEZ R, GEERAERTS D, STERCKX Y, et al, 2010. New toxins homologous to ParE belonging to three-component toxin-antitoxin systems in Escherichia coli O157:H7[J]. Molecular Microbiology, 76(3): 719-732.
[8]	HARMS A, LIESCH M, KÖRNER J, et al, 2017. A bacterial toxin-antitoxin module is the origin of inter-bacterial and inter-kingdom effectors of Bartonella[J]. PLoS Genetics, 13(10): e1007077.
[9]	HAU H H, GRALNICK J A, 2007. Ecology and biotechnology of the genus Shewanella[J]. Annual Review of Microbiology, 61(1): 237-258.
[10]	HEIDELBERG J F, PAULSEN I T, NELSON K E, et al, 2002. Genome sequence of the dissimilatory metal ion-reducing bacterium Shewanella oneidensis[J]. Nature Biotechnology, 20(11): 1118-1123.
[11]	JIANG YONG, POGLIANO J, HELINSKI D R, et al, 2002. ParE toxin encoded by the broad-host-range plasmid RK2 is an inhibitor of Escherichia coli gyrase[J]. Molecular Microbiology, 44(4): 971-979.
[12]	KONSTANTINIDIS K T, SERRES M H, ROMINE M F, et al, 2009. Comparative systems biology across an evolutionary gradient within the Shewanella genus[J]. Proceedings of the National Academy of Sciences of the United States of America, 106(37): 15909-15914.
[13]	LEPLAE R, GEERAERTS D, HALLEZ R, et al, 2011. Diversity of bacterial type II toxin-antitoxin systems: a comprehensive search and functional analysis of novel families[J]. Nucleic Acids Research, 39(13): 5513-5525.
[14]	LOVLEY D R, 2012. Electromicrobiology[J]. Annual Review of Microbiology, 66(1): 391-409.
[15]	MAGNUSON R D, 2007. Hypothetical functions of toxin-antitoxin systems[J]. Journal of Bacteriology, 189(17): 6089-6092.
[16]	MCKENZIE J L, ROBSON J, BERNEY M, et al, 2012. A VapBC toxin-antitoxin module is a posttranscriptional regulator of metabolic flux in mycobacteria[J]. Journal of Bacteriology, 194(9): 2189-2204.
[17]	NG C K, SIVAKUMAR K, LIU X, et al, 2013. Influence of outer membrane c-type cytochromes on particle size and activity of extracellular nanoparticles produced by Shewanella oneidensis[J]. Biotechnology and Bioengineering, 110(7): 1831-1837.
[18]	OGURA T, HIRAGA S, 1983. Mini-F plasmid genes that couple host cell division to plasmid proliferation[J]. Proceedings of the National Academy of Sciences of the United States of America, 80(15): 4784-4788.
[19]	PANDEY D P, GERDES K, 2005. Toxin-antitoxin loci are highly abundant in free-living but lost from host-associated prokaryotes[J]. Nucleic Acids Research, 33(3): 966-976.
[20]	ROBERTS R C, HELINSKI D R, 1992. Definition of a minimal plasmid stabilization system from the broad-host-range plasmid RK2[J]. Journal of Bacteriology, 174(24): 8119-8132.
[21]	SCHÄGGER H, 2006. Tricine-SDS-PAGE[J]. Nature Protocols, 1(1): 16-22.
[22]	SYED M A, LÉVESQUE C M, 2012. Chromosomal bacterial type II toxin-antitoxin systems[J]. Canadian Journal of Microbiology, 58(5): 553-562.
[23]	TSILIBARIS V, MAENHAUT-MICHEL G, MINE N, et al, 2007. What is the benefit to Escherichia coli of having multiple toxin-antitoxin systems in its genome?[J]. Journal of Bacteriology, 189(17): 6101-6108.
[24]	WANG XIAOXUE, KIM Y, HONG S H, et al, 2011. Antitoxin MqsA helps mediate the bacterial general stress response[J]. Nature Chemical Biology, 7(6): 359-366.
[25]	WEN YURONG, BEHIELS E, FELIX J, et al, 2014. The bacterial antitoxin HipB establishes a ternary complex with operator DNA and phosphorylated toxin HipA to regulate bacterial persistence[J]. Nucl Acids Research, 42(15): 10134-10147.
[26]	WIATROWSKI H A, WARD P M, BARKAY T, 2006. Novel reduction of mercury (II) by mercury-sensitive dissimilatory metal reducing bacteria[J]. Environmental Science and Technology, 40(21): 6690-6696.
[27]	WINTHER K S, GERDES K, 2011. Enteric virulence associated protein VapC inhibits translation by cleavage of initiator tRNA[J]. Proceedings of the National Academy of Sciences of the United States of America, 108(18): 7403-7407.
[28]	YAO JIANYUN, GUO YUNXUE, WANG PENGXIA, et al, 2018. Type II toxin/antitoxin system ParESO/CopASO stabilizes prophage CP4So in Shewanella oneidensis[J]. Environmental Microbiology, 20(3): 1224-1239.
[29]	YAO JIANYUN, GUO YUNXUE, ZENG ZHENSHUN, et al, 2015. Identification and characterization of a HEPN-MNT family type II toxin-antitoxin in Shewanella oneidensis[J]. Microbial Biotechnology, 8(6): 961-973.
[30]	YUAN JIE, STERCKX Y, MITCHENALL L A, et al, 2010. Vibrio cholerae ParE2 poisons DNA gyrase via a mechanism distinct from other gyrase inhibitors[J]. Journal of Biological Chemistry, 285(51): 40397-40408.

Shewanella oneidensis内源性大质粒上ParE-ParD家族毒素-抗毒素系统(SO_A0088-A0087)的鉴定

Characterization of a ParE-ParD toxin-antitoxin TA system (SO_A0087-A0088) on a megaplasmid of Shewanella oneidensis

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

图/表 5

参考文献 30

相关文章 1

Metrics

本文评价

推荐阅读 0