DNA interstrand crosslinking and sequence selectivity of dimethanesulphonates

Abstract

Members of the homologous series of alkanediol dimethanesulphonates of general formula H3C. S020. (CH2)n. 0. S02. CH3 have been tested for their ability to produce DNA interstrand crosslinking and DNA sequence selectivity of guanine-N7 alkylation. In a sensitive crosslinking gel assay the efficiency of DNA interstrand crosslink formation, dependent on the ability of the alkylating moiety to span critical nucleophilic distances within the DNA, was found at 6h to be 1, 6-hexanediol dimethanesulphonate (Hexa-DMS) (« = 6) > methylene dimethanesulphonate (MDMS) (n= 1)> 1, 8-octanediol dimethanesulphonate (Octa-DMS) (n = 8) > Busulphan (n = 4). The DNA interstrand crosslinking produced by MDMS was not due to either of its hydrolysis products, formaldehyde or methanesulphonic acid (MSA). In contrast the extent of monoalkyla tion at guanine-N7 as determined by a modified DNA sequencing technique was found to be Busul phan »Hexa-DMS = Octa-DMS, with a sequence selectivity somewhat less than that of other chemo therapeutic alkylating agents such as nitrogen mustards. MDMS at high levels induced a non-specific depurination as a result of the reduction in pH resulting from MSA release. More strikingly MDMS (and MSA) produced a single strong site of guanine reaction (depurination) in a guanine-rich 276 base pair fragment of pBR322 DNA in the sequence of 5'-ATGGTGG-3'. This was observed when non-specific depurination was negligible and was not seen with formic acid. Thus structurally similar alkylating agents can differ in their type and extent of DNA monoalkylation and interstrand crosslinking, and in some cases (e. g. MDMS/MSA) produce reactions with a high degree of selectivity. © Macmillan Press Ltd., 1991.