Structural Analysis of Virus Regulatory N6-Methyladenosine (m6A) Machinery of the Black Flying Fox ( Pteropus alecto ) and the Egyptian Fruit Bat ( Rousettus aegyptiacus ) Shows Evolutionary Conservation Amongst Mammals

N6-methyladenosine (m6A) is an abundant RNA epitranscriptomic modification in eukaryotes. The m6A machinery includes cellular writer, eraser and reader proteins that regulate m6A. ( ) (the Australian black flying fox) and ( ) (the Egyptian fruit bat) are bats associated with several viral zoonoses yet neglected in the field of m6A epigenetics studies. This study utilises various bioinformatics and in silico tools to genetically identify, characterise and annotate the m6A machinery in and . A range of bioinformatic tools were deployed to comprehensively characterise all known m6A-associated proteins of and Results: Phylogenetically, the m6A fat mass and obesity-associated protein (FTO) eraser placed the order Chiroptera (an order including all bat species) in a separate clade. Additionally, it showed the lowest identity matrices in and when compared to other mammals (74.1% and 72.8%) and (84.0% and 76.1%), respectively. When compared to humans, genetic loci-based analysis of and showed syntenic conservation in multiple flanking genes of 8 out the 10 m6A-associated genes. Furthermore, amino acid alignment and protein tertiary structure of the two bats' m6A machinery demonstrated conservation in the writers but not in erasers and readers, compared to humans. These studies provide foundational annotation and genetic characterisation of m6A machinery in two important species of bats which can be exploited to study bat-virus interactions at the interface of epitranscriptomics.