Chavez, R. M. | Search bioRxiv

bioRxiv2026

Rhomboid protease Rhbdl2 regulates macrophage recruitment and wound regeneration in zebrafish

Tissue regeneration requires tight control of immune cell behavior, yet the mechanisms that restrain immune-driven regenerative responses remain poorly defined. Here, we identify the rhomboid intramembrane serine protease Rhbdl2 as a critical regulator of regeneration in zebrafish. We generated rhbdl2 mutants by CRISPR Cas9 and found that it does not affect normal development, but triggers enhanced regenerative growth following injury, accompanied by increased macrophage accumulation at the wound site, which is accompanied by increased early apoptosis and proliferation. Proteomic analyses reveal increased Rac2 protein levels in rhbdl2 mutants, indicating dysregulated immune signaling. Functionally, Rac2 morpholino oligonucleotides mediated knockdown in rhbdl2 mutant larvae suppresses the elevated macrophage recruitment and enhanced tissue regenerative phenotype. Together, these findings uncover Rhbdl2 as an immune checkpoint that constrains macrophage driven enhanced regeneration, with vast implications for inflammatory disease, fibrosis, and tumor immune interactions.

immunology↗

bioRxiv2022

MSL2 targets histone genes in Drosophila virilis

Histone genes are amongst the most evolutionary conserved in eukaryotic genomes, yet cis-regulatory mechanisms of histone gene regulation differ considerably amongst species. In Drosophila melanogaster, an interaction between GA-rich cis elements in the H3/H4 promoter and the GA-binding transcription factor CLAMP is important for promoting histone gene regulation and factor recruitment to the locus. CLAMP also participates in male dosage compensation by recruiting the Male Specific Lethal Complex (MSLc) to the X-chromosome. We discovered that the male-specific protein of MSLc, MSL2, is recruited to the autosomal major histone locus in D. virilis but not to the minor locus or to the single histone locus in other species. While the histone coding sequences are well conserved between species, the critical GA-rich cis elements in the H3/H4 promoter are poorly conserved between D. melanogaster and D. virilis. We show that CLAMP still targets the two D. virilis histone loci in vivo. Further, CLAMP interacts with the D. virilis H3/H4 promoter in vitro, even when the poorly-conserved GA-rich cis elements are deleted, indicating that the protein interacts differently with the D. virilis promoter than it does with the D. melanogaster promoter. Since CLAMP and MSL2 directly interact in D. melanogaster, we propose that D. virilis CLAMP recruits MSL2 to an ectopic autosomal site through interaction with X-like cis elements. Further, localization of MSL2 to one of the D. virilis histone loci suggests that the loci are regulated differently and that males and females have different requirements for histone gene regulation.

genetics↗