Biochemical characterization of maintenance DNA methyltransferase DNMT-1 from silkworm, Bombyx mori.

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.

authors

Mitsudome T,Mon H,Xu J,Li Z,Lee JM,Patil AA,Masuda A,Iiyama K,Morokuma D,Kusakabe T

doi

10.1016/j.ibmb.2015.01.008

subject

Has Abstract

pub_date

2015-03-01 00:00:00

pages

55-65

eissn

0965-1748

issn

1879-0240

pii

S0965-1748(15)00015-6

journal_volume

58

pub_type

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