Analysis of the Tudor protein interactome in Drosophila germline development

Author

Ethan R. HackneyFollow

Abstract

Germ cells give rise to the egg and sperm cells and, therefore, to the next generation. In many organisms, germ cells assemble membraneless RNA-protein organelles called germ granules. Components of germ granules are required for germ cell specification. One of these crucial components is evolutionarily conserved Tudor domain-containing protein. Specifically, in the fruit fly Drosophila melanogaster this Tudor protein contains multiple Tudor domains. These domains are protein-protein interaction modules shown to interact with methylated amino acids of target proteins. We aim at understanding the molecular mechanisms of the germ granule assembly through detailed structure-function analysis of Tudor protein and its binding partners using Drosophila. Using genetic, imaging, and biochemical approaches, we found that in germ granules, Tudor associates with specific binding partner proteins including Piwi-protein Aubergine and ATP-dependent DEAD-Box RNA helicase eIF4A. Our data indicated that multiple Tudor domains are required for germline development and for interaction with different protein components of the granules. However, in addition to Tudor domains, we unexpectedly found that intrinsically disordered protein regions between Tudor domains are also involved in the binding with germ granule components. Our data suggest that Tudor recruits different proteins to the germ granules using its different regions for protein-protein interactions. Furthermore, we propose that Tudor complex assembly contributes to post-transcriptional gene regulation at germ granules.

Year manuscript completed

2025

Year degree awarded

2025

Author's Keywords

Tudor, Germ cells, membraneless granules, RNA-protein structures, Drosophila

Degree Awarded

Master of Science

Department

Biology

College/School

Jesse D. Jones College of Science, Engineering and Technology

Thesis Advisor

Alexey L Arkov

Committee Chair

Alexey L Arkov

Committee Member

Christopher Lennon

Committee Member

Dena Weinberger

Committee Member

Gary ZeRuth

Document Type

Thesis - Murray State Access only

Recommended Citation

Hackney, Ethan R., "Analysis of the Tudor protein interactome in Drosophila germline development" (2025). Murray State Theses and Dissertations. 382.
https://digitalcommons.murraystate.edu/etd/382