Heterologous Assembly of the Type VI Secretion System Empowers Laboratory Escherichia coli with Antimicrobial and Cell Penetration Capabilities

Cui, Yang1; Pei, Tong-Tong1; Liang, Xiaoye1; Li, Hao1; Zheng, Hao-Yu1; Dong, Tao1,2

2022-09-01

10.1128/aem.01305-22

APPLIED AND ENVIRONMENTAL MICROBIOLOGY   Impact Factor & Quartile

0099-2240

1098-5336

["The T6SS is a powerful and versatile protein delivery system. However, the complexity of its macromolecular structure and gene regulation makes it not a trivial task to reconstitute the T6SSs of pathogens in a nonpathogenic host.","The synthetic biology toolbox has amassed a vast number of diverse functional modules, but protein translocation modules for cell penetration and cytosol-to-cytosol delivery remain relatively scarce. The type VI secretion system (T6SS), commonly found in many Gram-negative pathogens, functions as a contractile device to translocate protein toxins to prokaryotic and eukaryotic cells. Here, we have assembled the T6SS of Aeromonas dhakensis, an opportunistic waterborne pathogen, in the common laboratory strain Escherichia coli BL21(DE3). We constructed a series of plasmids (pT6S) carrying the T6SS structural and effector genes under native or tetracycline-inducible promoters, the latter for controlled expression. Using fluorescence microscopy and biochemical analyses, we demonstrate a functional T6SS in E. coli capable of secreting proteins directly into the cytosol of neighboring bacteria and outcompeting a number of drug-resistant pathogens. The heterologous assembly of T6SS not only confers the lab workhorse E. coli with the cytosol-to-cytosol protein delivery capability but also demonstrates the potential for harnessing the T6SS of various pathogens for general protein delivery and antibacterial applications. IMPORTANCE The T6SS is a powerful and versatile protein delivery system. However, the complexity of its macromolecular structure and gene regulation makes it not a trivial task to reconstitute the T6SSs of pathogens in a nonpathogenic host. In this study, we have assembled an inducible T6SS in E. coli BL21(DE3) and demonstrated its functions in protein delivery and antimicrobial activities. The engineered T6SS empowers E. coli to deliver protein cargos into a wide range of prokaryotic and eukaryotic cells."]

secretion system T6SS synthetic biology interspecies interaction

SCI

English

Corresponding

National Key R&D Program of China[2020YFA0907200] ; National Natural Science Foundation of China[32030001]

Biotechnology & Applied Microbiology ; Microbiology

Biotechnology & Applied Microbiology ; Microbiology

WOS:000861346700001

AMER SOC MICROBIOLOGY

BIOLOGY & BIOCHEMISTRY

Web of Science

1.Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Joint Int Res Lab Metab & Dev Sci, Sch Life Sci & Biotechnol, Shanghai, Peoples R China
2.Southern Univ Sci & Technol, Sch Life Sci, Dept Immunol & Microbiol, Shenzhen, Peoples R China

Cui, Yang,Pei, Tong-Tong,Liang, Xiaoye,et al. Heterologous Assembly of the Type VI Secretion System Empowers Laboratory Escherichia coli with Antimicrobial and Cell Penetration Capabilities[J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY,2022.

Cui, Yang,Pei, Tong-Tong,Liang, Xiaoye,Li, Hao,Zheng, Hao-Yu,&Dong, Tao.(2022).Heterologous Assembly of the Type VI Secretion System Empowers Laboratory Escherichia coli with Antimicrobial and Cell Penetration Capabilities.APPLIED AND ENVIRONMENTAL MICROBIOLOGY.

Cui, Yang,et al."Heterologous Assembly of the Type VI Secretion System Empowers Laboratory Escherichia coli with Antimicrobial and Cell Penetration Capabilities".APPLIED AND ENVIRONMENTAL MICROBIOLOGY (2022).