Molecular and functional characterization of theDrosophila melanogasterconserved smORFome

Abstract

SummaryShort polypeptides encoded by small open reading frames (smORFs) are ubiquitously found in eukaryotic genomes and are important regulators of physiology, development, and mitochondrial processes. Here, we focus on a subset of 194 smORFs that are evolutionarily conserved between Drosophila melanogaster and humans. Many of these smORFs are conserved broadly in the bilaterian lineage, with ∼60 conserved in plants. Within these conserved smORFs, we observed remarkably heterogenous spatial and temporal expression patterns – indicating wide-spread tissue-specific and stage-specific mitochondrial architectures. In addition, an analysis of annotated functional domains revealed a predicted enrichment of smORF polypeptides localizing to mitochondria. We conducted an embryonic ribosome profiling experiment and applied a machine learning approach, finding support for translation of 82 of these smORFs during embryogenesis. We further embarked on functional characterization using CRISPR knockout/activation, RNAi knockdown, and cDNA overexpression, revealing diverse phenotypes. This study underscores the importance of identifying smORF function in disease and phenotypic diversity.

Two neuronal peptides encoded from a single transcript regulate mitochondrial complex III in Drosophila

“…Here, through an effort to systematically characterize human-conserved smORF genes in Drosophila ( Bosch et al, 2022 ), we identified two previously unstudied smORF peptides CG32736-PB and CG42308-PA that we named Sloth1 and Sloth2 based on their mutant phenotypes. Remarkably, both peptides are translated from the same transcript and share amino acid sequence similarity, suggesting that they encode paralogs.…”