Research output: Contribution to journal › Article › peer-review
Relative Efficiency of Recognition of 5-Methylcytosine and 5-Hydroxymethylcytosine by Methyl-Dependent DNA Endonuclease GlaI. / Petrova, D. V.; Naumenko, M. B.; Khantakova, D. V. et al.
In: Russian Journal of Bioorganic Chemistry, Vol. 45, No. 6, 01.11.2019, p. 625-629.Research output: Contribution to journal › Article › peer-review
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TY - JOUR
T1 - Relative Efficiency of Recognition of 5-Methylcytosine and 5-Hydroxymethylcytosine by Methyl-Dependent DNA Endonuclease GlaI
AU - Petrova, D. V.
AU - Naumenko, M. B.
AU - Khantakova, D. V.
AU - Grin, I. R.
AU - Zharkov, D. O.
PY - 2019/11/1
Y1 - 2019/11/1
N2 - Only a limited number of tools are available to study cytosine methylation in DNA. One of the representatives of the recently discovered methyl-dependent DNA endonucleases is an enzyme GlaI. It is of great interest for determining the methylation status of eukaryotic genomic DNA due to its ability to cleave only methylated DNA. However, the ability of the GlaI endonuclease to recognize oxidized derivatives of 5-methylcytosine (mC), in particular another epigenetic base, 5-hydroxymethylcytosine (hmC), has not yet been characterized. It is not possible to fully use the potential of GlaI in the analysis of methylation due to the notable occurrence of the latter in the DNA of mammals. In this study, the efficiency of cleavage of DNA substrates with various combinations of mC and hmC by methyl-dependent DNA-endonuclease GlaI was compared; the kinetic parameters of cleavage reactions for fully methylated and fully hydroxymethylated recognition site were determined. It was shown that in most cases GlaI recognized substrates containing mC better than substrates containing hmC in the same positions. The most effective hydrolysis of substrates containing modifications in the sequence 5'-GCGC-3'/3'-CGCG-5' required the presence of hmC not only in the central but also in the edge positions in both DNA chains as in the case of mC.
AB - Only a limited number of tools are available to study cytosine methylation in DNA. One of the representatives of the recently discovered methyl-dependent DNA endonucleases is an enzyme GlaI. It is of great interest for determining the methylation status of eukaryotic genomic DNA due to its ability to cleave only methylated DNA. However, the ability of the GlaI endonuclease to recognize oxidized derivatives of 5-methylcytosine (mC), in particular another epigenetic base, 5-hydroxymethylcytosine (hmC), has not yet been characterized. It is not possible to fully use the potential of GlaI in the analysis of methylation due to the notable occurrence of the latter in the DNA of mammals. In this study, the efficiency of cleavage of DNA substrates with various combinations of mC and hmC by methyl-dependent DNA-endonuclease GlaI was compared; the kinetic parameters of cleavage reactions for fully methylated and fully hydroxymethylated recognition site were determined. It was shown that in most cases GlaI recognized substrates containing mC better than substrates containing hmC in the same positions. The most effective hydrolysis of substrates containing modifications in the sequence 5'-GCGC-3'/3'-CGCG-5' required the presence of hmC not only in the central but also in the edge positions in both DNA chains as in the case of mC.
KW - 5-hydroxymethylcytosine
KW - 5-methylcytosine
KW - epigenetic methylation
KW - GlaI endonuclease
KW - EVOLUTION
KW - RESTRICTION
UR - http://www.scopus.com/inward/record.url?scp=85078621506&partnerID=8YFLogxK
U2 - 10.1134/S1068162019060323
DO - 10.1134/S1068162019060323
M3 - Article
AN - SCOPUS:85078621506
VL - 45
SP - 625
EP - 629
JO - Russian Journal of Bioorganic Chemistry
JF - Russian Journal of Bioorganic Chemistry
SN - 1068-1620
IS - 6
ER -
ID: 23577483