DNA-methyltransferase SsoII interaction with own promoter region binding site.
The investigation of Sso II DNA-methyltransferase (M.Sso II) interaction with the intergenic region of Sso II restriction-modification system was carried out. Seven guanine residues protected by M. Sso II from methylation with dimethylsulfate and thus probably involved in enzyme-DNA recognition were identified. Six of them are located symmetrically within the 15 bp inverted repeat inside the Sso II promoter region. The crosslinking of Sso II methyltransferase with DNA duplexes containing 5-bromo-2'-deoxyuridine (br5dU) instead of thymidine was performed. The crosslinked products were obtained in all cases, thus proving that tested thymines were in proximity with enzyme. The ability to produce the crosslinked products in one case was 2-5-fold higher than in other ones. This allowed us to imply that thymine residue in this position of the inverted repeat could be in contact with M. Sso II. Based on the experimental data, two symmetrical 4 bp clusters (GGAC), which could be involved in the interaction with M. Sso II in the DNA-protein complex, were identified. The model of M. Sso II interaction with its own promoter region was proposed.
Shilov
I
I
Institute of Agricultural Biotechnology, 42 Timiryazevskaya Street, Moscow 127550, Russia. ann@agrobio.msk.su
Tashlitsky
V
V
Khodoun
M
M
Vasil'ev
S
S
Alekseev
Y
Y
Kuzubov
A
A
Kubareva
E
E
Karyagina
A
A
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D015257:DNA-Cytosine Methylases / Q000235:genetics* / Q000378:metabolism*; D006147:Guanine; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D013941:Thymine
D006147:Guanine; C063974:DNA modification methylase SsoII; D015257:DNA-Cytosine Methylases; D013941:Thymine