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Cleavage by restriction endonucleases Mvai and EcoRIi of substrates modified in amino groups of heterocyclic bases

Kubareva, E.A.; Gromova, E.S.; Romanova, E.A.; Oretskaia, T.S.; Shabarova, Z.A.

Bioorganicheskaia Khimiia 16(4): 501-506

1990

ISSN/ISBN: 0132-3423
PMID: 2375778
Document Number: 360802
14-membered DNA-duplexes containing modified nucleoside residues, viz 4-N-methyldeoxycytidine (m4dC), 6-N-methyldeoxyadenosine (m6dA) or deoxyinosine (dI), in only one strand of the recognition site (CCA/TGG) of MvaI and EcoRII endonucleases were synthesized. It was shown that MvaI and EcoRII endonucleases interact with the exocyclic amino groups of the external dC residues and of the central dA residue of the recognition site exposed into the DNA major groove. These endonucleases which are isochizomers were found to possess different mechanisms of substrate cleavage. The ability of MvaI endonuclease to hydrolyze only unmodified strand of methylated duplexes allows one to make site-directed single-strand nicks in double-stranded DNA. Elimination of the 2-NH2-group located in the minor groove of DNA by substituting dI for dG had little, if any, effect on the hydrolytic activity of EcoRII and MvaI endonucleases.

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