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Bioscience Biotechnology and Biochemistry
Article . 2012 . Peer-reviewed
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Molecular Cloning, Expression, and Characterization of Starfish DNA (Cytosine-5)-methyltransferases

Authors: FUJIHARA, Yoshihito; MIYASAKO, Hiroshi; KATO, Kumiko; HAYASHI, Tadahiro; TORAYA, Tetsuo;

Molecular Cloning, Expression, and Characterization of Starfish DNA (Cytosine-5)-methyltransferases

Abstract

To determine whether and if so how a DNA methylation-dependent epigenetic mechanism for transcriptional gene silencing functions in Echinoderms, we cloned and sequenced dnmt1 and dnmt3 cDNAs of the starfish Asterina pectinifera. Since the Strongylocentrotus purpuratus genome has only two loci of DNA (cytosine-5)-methyltransferase genes encoding Dnmt1 and Dnmt3, they might constitute a sufficient set of dnmt genes in Echinoderms. The starfish Dnmt3 whose cDNA we cloned showed highest homology to a mammalian Dnmt3a2 splicing variant. Essentially all the characteristic motifs and sequences of the mammalian counterparts were found in the starfish Dnmts as well, except that a typical PCNA binding domain motif was lacking in the starfish Dnmt1. RT-PCR analysis indicated that the dnmt1 mRNA exists in both ovary and oocytes, but its levels in other tissues were very low or almost negligible. In contrast, the dnmt3 mRNA was detected only in the ovary, and not at all in the oocytes. The size of a dnmt1 transcript was about 6.5 kb on Northern blot analysis. On heterologous expression, the starfish Dnmt1 protein was expressed in insect cells in catalytically active form.

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Keywords

Recombinant Fusion Proteins, Amino Acid Motifs, Molecular Sequence Data, DNA methyltransferase, starfish, Escherichia coli, Sf9 Cells, Animals, DNA (Cytosine-5-)-Methyltransferases, Cloning, Molecular, Strongylocentrotus purpuratus, Phylogeny, Gene Library, Sequence Homology, Amino Acid, Ovary, Protein Structure, Tertiary, Isoenzymes, Organ Specificity, Asterina, Dnmt1, Oocytes, cDNA cloning, Dnmt3, Female, Sequence Alignment

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
5
Average
Average
Average
bronze