Most populations of the aphid Myzus persicae have amplified genes (up to 80 copies) encoding the insecticide-detoxifying esterase E4. This paper reports the analysis of methylation of the E4 gene and its flanking DNA with the use of methylation-sensitive restriction enzymes, CpG profiling and bisulphite sequencing. In combination these show that E4 has 5-methylcytosine confined to CpG doublets, as previously shown for vertebrate genomes; this is the first such report for an insect gene. The methylation is present within the gene but absent from upstream regions, including the 5´ CpG-rich region around the start of transcription, and from 3´ flanking DNA. Methylated E4 genes are expressed; loss of the 5-methylcytosine is correlated with a loss of transcription, although this is not accompanied by a global loss of the 5-methylcytosine present in the aphid genome. These results suggest that the methylation of E4 has a positive role in expression, and call into question the widely held view that methylation in invertebrate genomes is confined to regions that do not contain genes and that methylation is always associated with gene silencing.
- DNA methylation
- gene amplification
- gene expression
- The Biochemical Society, London © 2000