The C-terminal ZZ domain of the Drosophila ORB2 RNA-binding protein is required for spermatid individualization

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Abstract

ORB2 is the Drosophila ortholog of the human CPEB2-4 family of RNA-binding proteins, which include a conserved C-terminal Zinc-binding (‘ZZ’) domain. We have recently shown that this domain interacts with several translation co-repressors in the early embryo, and that deletion of this domain from the endogenous orb2 gene results in derepression of its target mRNAs (LOW et al . 2025). Here we assess the effect of deletion of the ZZ domain on spermatogenesis. We find that ORB2ΔZZ protein is no longer localized to the distal tip of spermatids; that in orb2 ΔZZ testes, additional proteins known to play a role in spermatid individualization – ORB, IMP, SOTI – are mislocalized; that the SOTI-dependent Cleaved Caspase 3 gradient no longer forms; that individualization complexes are defective; and that orb2 ΔZZ flies are sterile and lack mature sperm.

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