Decoding the molecular principles underlying tumor-suppressive or oncogenic properties of large non-coding RNAs

• Applicant: Dr. Mathias Munschauer
• Subject Area: General Genetics and Functional Genome Biology ; Biochemistry ; Cell Biology
• Term: from 2015 to 2018
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 299175676

Decoding the molecular principles underlying tumor-suppressive or oncogenic properties of large non-coding RNAs. One of the most intriguing questions in current genome biology concerns the functional role of large non-coding RNAs (lncRNAs). LncRNAs show remarkable tissue specific expression, clear patterns of conservation and gene-like chromatin signatures. Importantly, individual lncRNAs are emerging as powerful regulators of tumorigenic events and can thus be classified as tumor suppressors or oncogenes. In contrast to their regulatory importance, mechanistic understanding of their oncogenic or tumor-suppressive properties is lacking. A key prerequisite to reveal the molecular principles underlying lncRNA function is the detailed knowledge of their protein-binding partners. Here, I utilize RNA antisense purification followed by quantitative mass spectrometry to identify the direct protein-binding partners of two tumor-suppressive and two oncogenic lncRNAs. Following identification of specific protein binders, I will use crosslinking and immunoprecipitation to map their RNA binding sites. Harnessing this information, I will systematically classify captured protein interactions into functionally essential and non-essential interactions. I will mutate distinct protein binding domains within lncRNAs and monitor known lncRNA-driven changes in the tumorigenic potency of respective cells. Thus, I will be able to connect lncRNA-dependent tumorigenic phenotypes to the specific recruitment of a distinct set of protein regulators to each lncRNA. This research will illuminate the molecular components that mediate oncogenic or tumor-suppressive lncRNA properties and further reveal important design principles of these regulators. Exploiting these insights, I will build synthetic lncRNA replacement constructs based on functionally important protein interaction domains and test if these constructs retain desired regulatory activities (e.g. tumor-suppressive activity). I envision that this approach will open the door for novel RNA-based therapeutic strategies and transform our understanding of lncRNA-mediated gene regulation in cancer biology.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Professor Dr. Eric Lander