Molecular cloning and expression-profile analysis of sea cucumber DNA (Cytosine-5)-methyltransferase 1 and methyl-CpG binding domain type 2/3 genes during aestivation

摘要

The sea cucumber Apostichopus japonicus Selenka survives high summer temperature by entering aestivation, characterized by hypometabolism and global gene silencing. We investigated the hypothesis that aestivation is associated with DNA methylation-dependent epigenetic mechanisms by cloning, sequencing and measuring the transcript abundances of two genes dnmt1 and mbd2/3, which comprise the DNA methylation system in A. japonicus Selenka. The deduced amino acid sequences and characteristic motifs of sea cucumber DNMT1 and MBD2/3 showed high homology to those of their mammalian counterparts. Quantitative real-time RT-PCR analysis showed that dnmt1 and mbd2/3 genes were similarly expressed in all four tissues examined (intestine, respiratory tree, muscle and body wall). Dnmt1 expression in the intestine was up-regulated during deep aestivation (P< 0.05), while mbd2/3 was over-expressed in both the intestine and respiratory tree during the same period (P< 0.01). No differences in expression levels were observed between other tissues. The results of this study suggest that DNA methylation may be involved in transcriptional silencing, and that the intestine is the major site for epigenetic regulation during aestivation in the sea cucumber.