Argonaute protein regulation by N-terminal intrinsically disordered regions

• Applicant: Professor René Ketting, Ph.D.
• Subject Area: Biochemistry ; Cell Biology
• Term: since 2025
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 569879053

Intrinsically disordered regions (IDRs) of proteins are prevalent. One context in which these are heavily studied is subcellular organization through phase separation. However, such IDRs may also have other functions. We study the function of Argonaute proteins in the process of RNA interference, and how this can be inherited across generations, using the nematode C. elegans as an important model organism. These Argonaute proteins often contain N-terminal IDRs, and they are often found in phase separated structures, known as germ granules. Yet, our preliminary work suggests that N-terminal IDRs of Argonaute proteins do not strongly affect their localization to these granules, but that they do affect Argonaute function. One way how this may work is by affecting the loading of the Argonaute protein. This project proposes experiments that will dissect the function of N-terminal IDRs of two specific Argonaute proteins, WAGO-1 and WAGO-3. This will include studies on enzymes that proteolytically process these IDRs, studies that address the loading, stability and subcellular localization of Argonaute proteins and studies on the inheritance of RNAi across generations. Coupled to these studies will be biochemical approaches to identify novel players in the studied mechanisms. Finally, the RNAi field in nematodes has been driving forward our understanding of this process. Yet, we mostly lack structural insights. In collaboration, we aim to get first structures of Argonaute proteins from C. elegans. Combined, these studies will break new ground in our understanding of Argonaute protein function. As Argonaute proteins are strongly conserved across species, we expect that the envisioned new insights will help propel Argonaute-related work in other species as well. Finally, these studies will help to establish IDRs as much more than ‘just’ being involved in phase separation.

• DFG Programme: Research Grants