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The replication focus targeting sequence (RFTS) domain is a DNA-competitive inhibitor of Dnmt1
Journal article   Open access   Peer reviewed

The replication focus targeting sequence (RFTS) domain is a DNA-competitive inhibitor of Dnmt1

Farisa Syeda, Rebecca L Fagan, Matthew Wean, George V Avvakumov, John R Walker, Sheng Xue, Sirano Dhe-Paganon and Charles Brenner
The Journal of biological chemistry, Vol.286(17), pp.15344-15351
04/29/2011
DOI: 10.1074/jbc.M110.209882
PMCID: PMC3083197
PMID: 21389349
url
https://doi.org/10.1074/jbc.M110.209882View
Published (Version of record) Open Access

Abstract

Dnmt1 (DNA methyltransferase 1) is the principal enzyme responsible for maintenance of cytosine methylation at CpG dinucleotides in the mammalian genome. The N-terminal replication focus targeting sequence (RFTS) domain of Dnmt1 has been implicated in subcellular localization, protein association, and catalytic function. However, progress in understanding its function has been limited by the lack of assays for and a structure of this domain. Here, we show that the naked DNA- and polynucleosome-binding activities of Dnmt1 are inhibited by the RFTS domain, which functions by virtue of binding the catalytic domain to the exclusion of DNA. Kinetic analysis with a fluorogenic DNA substrate established the RFTS domain as a 600-fold inhibitor of Dnmt1 enzymatic activity. The crystal structure of the RFTS domain reveals a novel fold and supports a mechanism in which an RFTS-targeted Dnmt1-binding protein, such as Uhrf1, may activate Dnmt1 for DNA binding.
 DNA Replication Protein Structure, Tertiary DNA (Cytosine-5-)-Methyltransferase 1 Humans Nucleosomes - metabolism Crystallography, X-Ray DNA - metabolism DNA (Cytosine-5-)-Methyltransferases - antagonists & inhibitors DNA (Cytosine-5-)-Methyltransferases - chemistry Animals Chickens Kinetics DNA-Binding Proteins

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