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

FEBS Lett. 2023 Jul;597(14):1848-1867. doi: 10.1002/1873-3468.14668. Epub 2023 Jun 8.

Abstract

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.

Keywords: Drosophila; Me31B; RNA helicase; germ granule; germline; oogenesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism
  • Drosophila Proteins* / metabolism
  • Drosophila* / genetics
  • Drosophila* / metabolism
  • Germ Cells / metabolism
  • Mutation

Substances

  • Drosophila Proteins
  • DEAD-box RNA Helicases
  • Me31B protein, Drosophila