Deciphering metatranslatomes - the chain link to understand functionality of gut microbiota

• Applicant: Privatdozent Dr. Klaus Neuhaus
• Subject Area: Nutritional Sciences
• Term: from 2017 to 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 327608356

Ribosomal footprinting (Ribo-seq) is a recently developed technique, which allows detection of mRNA under translation, thus providing a chain link between transcriptome and proteome data. For instance, we could show that translatome data acquired by Ribo-seq correlate well to proteomic data. Thus, Ribo-seq allows mapping of metabolic reactions to proteomic changes in cells. In addition, transcriptional and translational regulation can be elucidated and differentiation between non-coding and coding RNAs is possible. All of this results in unprecedented chances to understand microbial regulatory networks of genes and proteins at the same time. Ribo-seq has been used currently on single bacteria only, but mixed populations are the rule in nature. The intestinal microbiota, including its functionality, has important implications on the host, in terms of illness in case of dysbiosis. However, todays microbiota research is primarily focused on the study of eco-system diversity and composition. Certain metabolic changes are correlated to species abundance but the link often remains vague. In the present project, we will implement Ribo-seq for metatrans-latome analysis of complex microbial communities. Charting of the data to metabolic network maps allows assignment of microbial activities, either in functional microbioal groups or individual species. From an experimental point of view, Ribo-seq reduces sample complexity because the technique is inherently blind for nucleic acids not actively engaged. Likewise, since only translation of the microbiota is observed, the method is blind to otherwise interfering proteins of foreign sources, i.e., originating from the host or food. The model chosen will be IBD-susceptible IL-10 knock-out mice in combination with an established and simplified microbial gut community. The novel data will provide the chain link between sole bacterial abundance and microbiota functionality in this mouse model, paving way for future human studies.

• DFG Programme: Research Grants