RRC ID 33137
Author Mitsudome T, Mon H, Xu J, Li Z, Lee JM, Patil AA, Masuda A, Iiyama K, Morokuma D, Kusakabe T.
Title Biochemical characterization of maintenance DNA methyltransferase DNMT-1 from silkworm, Bombyx mori.
Journal Insect Biochem. Mol. Biol.
Abstract DNA methylation is an important epigenetic mechanism involved in gene expression of vertebrates and invertebrates. In general, DNA methylation profile is established by de novo DNA methyltransferases (DNMT-3A, -3B) and maintainance DNA methyltransferase (DNMT-1). DNMT-1 has a strong substrate preference for hemimethylated DNA over the unmethylated one. Because the silkworm genome lacks an apparent homologue of de novo DNMT, it is still unclear that how silkworm chromosome establishes and maintains its DNA methylation profile. As the first step to unravel this enigma, we purified recombinant BmDNMT-1 using baculovirus expression system and characterized its DNA-binding and DNA methylation activity. We found that the BmDNMT-1 preferentially methylates hemimethylated DNA despite binding to both unmethylated and hemimethylated DNA. Interestingly, BmDNMT-1 formed a complex with DNA in the presence or absence of methyl group donor, S-Adenosylmethionine (AdoMet) and the AdoMet-dependent complex formation was facilitated by Zn(2+) and Mn(2+). Our results provide clear evidence that BmDNMT-1 retained the function as maintenance DNMT but its sensitivity to metal ions is different from mammalian DNMT-1.
Volume 58
Pages 55-65
Published 2015-3
DOI 10.1016/j.ibmb.2015.01.008
PII S0965-1748(15)00015-6
PMID 25623240
MeSH Amino Acid Sequence Animals Baculoviridae / metabolism Bombyx / enzymology Bombyx / genetics Bombyx / metabolism* Cell Line DNA / metabolism DNA (Cytosine-5-)-Methyltransferases / chemistry* DNA (Cytosine-5-)-Methyltransferases / genetics* DNA (Cytosine-5-)-Methyltransferases / metabolism DNA Methylation Dimerization Larva / enzymology Larva / genetics Larva / metabolism Recombinant Proteins Sequence Alignment
IF 3.562
Times Cited 3
WOS Category ENTOMOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Silkworms p50