Time-Resolved Proteome Analysis of Listeria monocytogenes during Infection Reveals the Role of the AAA+ Chaperone ClpC for Host Cell Adaptation

被引:6
作者
Birk, Marlene S. [1 ]
Ahmed-Begrich, Rina [1 ]
Tran, Stefan [1 ,2 ]
Elsholz, Alexander K. W. [1 ]
Frese, Christian K. [1 ]
Charpentier, Emmanuelle [1 ]
机构
[1] Max Planck Unit Sci Pathogens, Berlin, Germany
[2] Max Planck Inst Mol Biomed, Munster, Germany
关键词
proteomics; tandem-mass-tag; pulsed SILAC; Listeria monocytogenes; proteolysis; ClpC; AAA+; infection; adaptation; host-pathogen interactions; mass spectrometry; TANDEM MASS TAGS; STAPHYLOCOCCUS-AUREUS; DEPENDENT PROTEOLYSIS; FLAGELLAR MOTILITY; STRESS TOLERANCE; VIRULENCE; BACILLUS; METABOLISM; PROTEINS; MUTANT;
D O I
10.1128/mSystems.00215-21
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The cellular proteome comprises all proteins expressed at a given time and defines an organism's phenotype under specific growth conditions. The proteome is shaped and remodeled by both protein synthesis and protein degradation. Here, we developed a new method which combines metabolic and chemical isobaric peptide labeling to simultaneously determine the time-resolved protein decay and de novo synthesis in an intracellular human pathogen. We showcase this method by investigating the Listeria monocytogenes proteome in the presence and absence of the AAA+ chaperone protein ClpC. ClpC associates with the peptidase ClpP to form an ATP-dependent protease complex and has been shown to play a role in virulence development in L. monocytogenes. However, the mechanism by which ClpC is involved in the survival and proliferation of intracellular L. monocytogenes remains elusive. Employing this new method, we observed extensive proteome remodeling in L. monocytogenes upon interaction with the host, supporting the hypothesis that ClpC-dependent protein degradation is required to initiate bacterial adaptation mechanisms. We identified more than 100 putative ClpC target proteins through their stabilization in a clpC deletion strain. Beyond the identification of direct targets, we also observed indirect effects of the clpC deletion on the protein abundance in diverse cellular and metabolic pathways, such as iron acquisition and flagellar assembly. Overall, our data highlight the crucial role of ClpC for L. monocytogenes adaptation to the host environment through proteome remodeling. IMPORTANCE Survival and proliferation of pathogenic bacteria inside the host depend on their ability to adapt to the changing environment. Profiling the underlying changes on the bacterial proteome level during the infection process is important to gain a better understanding of the pathogenesis and the host-dependent adaptation processes. The cellular protein abundance is governed by the interplay between protein synthesis and decay. The direct readout of these events during infection can be accomplished using pulsed stable-isotope labeling by amino acids in cell culture (SILAC). Combining this approach with tandemmass-tag (TMT) labeling enabled multiplexed and time-resolved bacterial proteome quantification during infection. Here, we applied this integrated approach to investigate protein turnover during the temporal progression of adaptation of the human pathogen L. monocytogenes to its host on a system-wide scale. Our experimental approach can easily be transferred to probe the proteome remodeling in other bacteria under a variety of perturbations.
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页数:19
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