The amino-terminus of mouse DNA methyltransferase 1 forms an independent domain and binds to DNA with the sequence involving PCNA binding motif.
DNA methylation patterns in genome are maintained during replication by a DNA methyltransferase Dnmt1. Mouse Dnmt1 is a 180 kDa protein comprising the N-terminal regulatory domain, which covers 2/3 of the molecule, and the rest C-terminal catalytic domain. In the present study, we demonstrated that the limited digestion of full-length Dnmt1 with different proteases produced a common N-terminal fragment, which migrated along with Dnmt1 (1-248) in SDS-polyacrylamide gel electrophoresis. Digestion of the N-terminal domains larger than Dnmt1 (1-248) with chymotrypsin again produced the fragment identical to the size of Dnmt1 (1-248). These results indicate that the N-terminal domain of 1-248 forms an independent domain. This N-terminal domain showed DNA binding activity, and the responsible sequence was narrowed to the 79 amino acid residues involving the proliferating cell nuclear antigen (PCNA) binding motif. The DNA binding activity did not distinguish between DNA methylated and non-methylated states, but preferred to bind to the minor groove of AT-rich sequence. The DNA binding activity of the N-terminal domain competed with the PCNA binding. We propose that DNA binding activity of the N-terminal domain contributes to the localization of Dnmt1 to AT-rich sequence such as Line 1, satellite, and the promoter of tissue-specific silent genes.
Suetake
Isao
I
Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871. suetake@protein.osaka-u.ac.jp
Hayata
Daichika
D
Tajima
Shoji
S
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain / Q000235:genetics*; D004247:DNA / Q000378:metabolism*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000737:chemistry* / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004214:Distamycins / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D008739:Methyl Green / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D009429:Netropsin / Q000494:pharmacology; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics*; D011485:Protein Binding / Q000187:drug effects; D016415:Sequence Alignment
D004268:DNA-Binding Proteins; D004214:Distamycins; D007211:Indoles; D018809:Proliferating Cell Nuclear Antigen; C007293:DAPI; D009429:Netropsin; C030000:stallimycin; D008739:Methyl Green; D004247:DNA; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619381:Dnmt1 protein, mouse