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MMS–derived DSBs and SMD require abasic site (AP) intermediates and are prevented by a mag1 deletion mutation or methoxyamine (MX).
The logarithmically growing apn1/2 and apn1/2 mag1 cells in YPDA were arrested at G2/M by nocodazole, treated with MMS (0.1%, 15 min) in PBS and returned to YPDA+nocodazole and incubated further. The cells were incubated with MX (100 mM) prior to, during and following MMS treatment. Cells were collected at the indicated times and processed for PFGE analysis.