Secretome and Comparative Proteomics of Yersinia pestis Identify Two Novel E3 Ubiquitin Ligases That Contribute to Plague Virulence
| Autor: | Yuling Chen, Songbiao Zhu, Tong Wang, Yajun Song, Shiyang Cao, Haiteng Deng, Yanfeng Yan, Yafang Tan, Ruifu Yang, Zongmin Du |
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| Rok vydání: | 2021 |
| Předmět: |
Proteomics
Yersinia pestis Ubiquitin-Protein Ligases Fv flea vector Virulence E3 ubiquitin ligases Biochemistry Analytical Chemistry Type three secretion system Microbiology comparative proteomics 03 medical and health sciences Bacterial Proteins PRM parallel reaction monitoring NEL novel E3 ubiquitin ligase Animals Humans T3SS type III secretion system Molecular Biology Secretome 030304 developmental biology OMV outer membrane vesicle Mice Inbred BALB C Plague Mh mammalian host 0303 health sciences biology AGC automatic gain control Research Yops Yersinia outer proteins 030302 biochemistry & molecular biology TMT tandem mass tag biology.organism_classification PD Proteome Discoverer Ubiquitin ligase Secretory protein Membrane protein Mutation NCE normalized collisional energy Proteome biology.protein Female HeLa Cells |
| Zdroj: | Molecular & Cellular Proteomics : MCP |
| ISSN: | 1535-9476 |
| DOI: | 10.1016/j.mcpro.2021.100066 |
| Popis: | Plague is a zoonotic disease that primarily infects rodents via fleabite. Transmission from flea to host niches requires rapid adaption of Yersinia pestis to the outer environments to establish infection. Here, quantitative proteome and secretome analyses of Y. pestis grown under conditions mimicking the two typical niches, i.e., the mammalian host (Mh) and the flea vector (Fv), were performed to understand the adaption strategies of this deadly pathogen. A secretome of Y. pestis containing 308 proteins has been identified using TMT-labeling mass spectrometry analysis. Although some proteins are known to be secreted, such as the type III secretion substrates, PsaA and F1 antigen, most of them were found to be secretory proteins for the first time. Comparative proteomic analysis showed that membrane proteins, chaperonins and stress response proteins are significantly upregulated under the Mh condition, among which the previously uncharacterized proteins YP_3416∼YP_3418 are remarkable because they cannot only be secreted but also translocated into HeLa cells by Y. pestis. We further demonstrated that the purified YP_3416 and YP_3418 exhibited E3 ubiquitin ligase activity in in vitro ubiquitination assay and yp_3416∼3418 deletion mutant of Y. pestis showed significant virulence attenuation in mice. Taken together, our results represent the first Y. pestis secretome, which will promote the better understanding of Y. pestis pathogenesis, as well as the development of new strategies for treatment and prevention of plague. Graphical Abstract Highlights • Quantitative proteome analysis of Y. pestis was performed to understand its adaption strategies. • A secretome of Y. pestis containing 308 proteins has been identified. • YP_3416 and YP_3418 exhibited strong E3 ubiquitin ligase activity. • yp_3416∼3418 deletion mutant of Y. pestis showed significant virulence attenuation in mice. In Brief Rapid adaption to the environments is critical for microbes to establish infection. Quantitative proteome and secretome analyses of Y. pestis grown under conditions mimicking its two typical natural niches were performed to understand the adaption strategies of this deadly pathogen. We identified three secreted proteins that can be translocated into host cells and further demonstrated that two of them show strong E3 ubiquitin ligase activity and contribute significantly to the virulence of Y. pestis. |
| Databáze: | OpenAIRE |
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