View Program - 63rd Annual Drosophila Research Conference

Drosophila Me31B (DDX6) is a conserved ATP-dependent RNA helicase that plays role in post-transcriptional RNA regulation to ensure the correct spatial and temporal expression of germline mRNAs. This, in turn, is crucial for the proper germline development in many animals. However, Me31B’s role and molecular mechanism are not clear. To study this, we aim to first understand the functions of important domains/motifs of the protein. Therefore, we used CRISPR gene-editing technique and generated Drosophila strains mutant for functionally important motifs in Me31B including the two RecA-like domains, N-terminal domain and C-terminal domain, motifs involved in ATPase and helicase activities such as DEAD-box motif and HRIGR motif, motifs that cause human developmental defects if mutated such as QxxR motif, and protein-binding motifs such as FDF-pocket motif and W-pocket motif. Further, we are conducting a series of phenotype characterization experiments on the mutants including quantification of the expressed mutant proteins, confocal microscopic examination of the mutant proteins’ localization in the ovaries, and fertility assays of the mutant strains. Considering that Me31B/DDX6 family proteins are also expressed in certain soma like neurons and other organisms such as yeast, worm, mouse, and human, the results from this study could further help us understand Me31B/DDX6 family proteins’ likely conserved roles in different cell types and species.

Analysis of RNA Helicase Me31B’s Molecular Mechanism in Germline Development by Motif Mutations