4.5 Article

Mutational analysis of the functional motifs of the DEAD-box RNA helicase Me31B/DDX6 in Drosophila germline development

期刊

FEBS LETTERS
卷 597, 期 14, 页码 1848-1867

出版社

WILEY
DOI: 10.1002/1873-3468.14668

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Drosophila; germ granule; germline; Me31B; oogenesis; RNA helicase

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In this study, the in vivo functions of key motifs/domains of Me31B were investigated using the Drosophila germline as a model. Mutations in different domains were generated using CRISPR and the effects on various processes of germline development were characterized. The results provide insights into the working mechanism of Me31B and its importance in germline regulation.
Me31B/DDX6 is a DEAD-box family RNA helicase playing roles in post-transcriptional RNA regulation in different cell types and species. Despite the known motifs/domains of Me31B, the in vivo functions of the motifs remain unclear. Here, we used the Drosophila germline as a model and used CRISPR to mutate the key Me31B motifs/domains: helicase domain, N-terminal domain, C-terminal domain and FDF-binding motif. Then, we performed screening characterization on the mutants and report the effects of the mutations on the Drosophila germline, on processes such as fertility, oogenesis, embryo patterning, germline mRNA regulation and Me31B protein expression. The study indicates that the Me31B motifs contribute different functions to the protein and are needed for proper germline development, providing insights into the in vivo working mechanism of the helicase.

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