Abstract
BackgroundThe phylum Platyhelminthes (flatworms) contains an important group of bilaterian organisms responsible for many debilitating and chronic infectious diseases of human and animal populations inhabiting the planet today. In addition to their biomedical and veterinary relevance, some platyhelminths are also frequently used models for understanding tissue regeneration and stem cell biology. Therefore, the molecular (genetic and epigenetic) characteristics that underlie trophic specialism, pathogenicity or developmental maturation are likely to be pivotal in our continued studies of this important metazoan group. Indeed, in contrast to earlier studies that failed to detect evidence of cytosine or adenine methylation in parasitic flatworm taxa, our laboratory has recently defined a critical role for cytosine methylation in Schistosoma mansoni oviposition, egg maturation and ovarian development. Thus, in order to identify whether this epigenetic modification features in other platyhelminth species or is a novelty of S. mansoni, we conducted a study simultaneously surveying for DNA methylation machinery components and DNA methylation marks throughout the phylum using both parasitic and non-parasitic representatives.ResultsFirstly, using both S. mansoni DNA methyltransferase 2 (SmDNMT2) and methyl-CpG binding domain protein (SmMBD) as query sequences, we illustrate that essential DNA methylation machinery components are well conserved throughout the phylum. Secondly, using both molecular (methylation specific amplification polymorphism, MSAP) and immunological (enzyme-linked immunoabsorbent assay, ELISA) methodologies, we demonstrate that representative species (Echinococcus multilocularis, Protopolystoma xenopodis, Schistosoma haematobium, Schistosoma japonicum, Fasciola hepatica and Polycelis nigra) within all four platyhelminth classes (Cestoda, Monogenea, Trematoda and ‘Turbellaria’) contain methylated cytosines within their genome compartments.ConclusionsCollectively, these findings provide the first direct evidence for a functionally conserved and enzymatically active DNA methylation system throughout the Platyhelminthes. Defining how this epigenetic feature shapes phenotypic diversity and development within the phylum represents an exciting new area of metazoan biology.
Highlights
The phylum Platyhelminthes contains an important group of bilaterian organisms responsible for many debilitating and chronic infectious diseases of human and animal populations inhabiting the planet today
For the first time, that DNA methylation and essential DNA methylation machinery components (DNMTs and Methyl-CpG binding domain (MBD)) are present in both parasitic and free-living flatworm species
DNA methylation is found in all four platyhelminth classes Having established that representative species across all four classes within the Platyhelminthes contain a S. mansoni DNA methyltransferase 2 (SmDNMT2) homolog (Figures 1 and 2), we investigated whether DNA methylation was conserved across the phylum (Figure 5)
Summary
The phylum Platyhelminthes (flatworms) contains an important group of bilaterian organisms responsible for many debilitating and chronic infectious diseases of human and animal populations inhabiting the planet today. In addition to their biomedical and veterinary relevance, some platyhelminths are frequently used models for understanding tissue regeneration and stem cell biology. Metazoan DNA methyltransferases (DNMT1, DNMT2, DNMT3a/3b [7]) catalyse this feature by transferring a methyl group (CH3) from S-adenosylmethionine (SAM) to the 5-carbon (C5) position of cytosine in gDNA These ‘epigenetic marks’ are subsequently recognised by methyl-CpG binding domain proteins (MeCP2 and MBD1-4) and converted into signals necessary for generating phenotypic diversity [8]. DNMTs and MBDs (complexed with other proteins [9]) comprise the core metazoan DNA methylation system found in both vertebrate and invertebrate species
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