Genome-wide mapping of the Galleria mellonella transcription start sites during fungal infection and treatment.

Galleria mellonella is an emerging experimental model, but its current genome annotation lacks transcription start site (TSS) information. Because TSS annotation is essential for understanding transcriptional regulation, we used low quantity single strand CAGE (LQ-ssCAGE) to map cap analysis of gene expression (CAGE)-defined TSSs from 72 G. mellonella larval samples. Among these, 36 larvae were infected with the fungal pathogen Madurella mycetomatis and 36 served as healthy controls, allowing identification of TSSs under both infected and healthy conditions. Infected and healthy larvae were treated with itraconazole or ravuconazole. To assess key characteristics of the CAGE data, TSS peaks and putative enhancers were intersected with the existing genome annotation for annotation and characterization. This analysis identified two sets of TSS peaks, permissive and robust, representing the first draft TSS atlas for G. mellonella. The atlas-derived annotations showed a markedly stronger core promoter sequence signature than the NCBI reference annotation, supporting improved resolution and accuracy of TSS annotation in the atlas.