A Proteogenomic Resource Enabling Integrated Analysis of Listeria Genotype–Proteotype–Phenotype Relationships

• DOI: 10.1021/acs.jproteome.9b00842
• Publication Date: 2020-02-24T21:45:26Z
• Authors: Adithi R. Varadar...; Sandra Goetze; Maria P. Pavlou; Virginie Grosboillot; Yang Shen; Martin J. Loessner; Christian H. Ahrens; Bernd Wollscheid
• Keywords: Proteogenomics

Listeria monocytogenes is an opportunistic foodborne pathogen responsible for listeriosis, a potentially fatal disease. Many different strains and serotypes exist, but proteogenomic resource that bridges the gap in our molecular understanding of relationships between genotypes phenotypes via proteotypes still missing. Here, we devised next-generation proteogenomics strategy enables community to rapidly proteotype relate this information back genotype. Based on sequencing de novo assembly two most commonly used model strains, EGD-e ScottA, established comprehensive databases. A genome comparison core- strain-specific genes virulence differences. Next, DIA/SWATH-based proteotyping strategy, including new robust sample preparation workflow, reproducible, sensitive, relative quantitative measurement proteotypes. This reusable publicly available DIA/SWATH library covers 70% open reading frames represents extensive spectral analysis date. We these resources investigate states mimicking upper gastrointestinal passage. Exposure bile salts at 37 °C, which simulates conditions encountered duodenum, showed significant perturbations increase FlaA, structural protein flagella. Given known lose its flagella above 30 was unexpected finding. The formation flagella, might have implications infectivity, validated by parallel reaction monitoring light scanning electron microscopy. flaA transcript levels did not change significantly upon exposure suggesting regulation post-transcriptional level. Together, analyses provide toolbox enabling genotype-proteotype-phenotype relationships.