Sequence context effect for hMSH2-hMSH6 mismatch-dependent activation

Numerous DNA mismatches and lesions activate MutS homologue (MSH) ATPase activity that is essential for mismatch repair (MMR). We have found that a mismatch embedded in a nearest-neighbor sequence context containing symmetric 3′-purines (2 × 3′-purines) enhanced, whereas symmetric 3′-pyrimidines (2 × 3′-pyrimidines) reduced, hMSH2-hMSH6 ATPase activation. The 3′-purine/pyrimidine effect was most evident for G-containing mispairs. A similar trend pervaded mismatch binding (KD) and the melting of unbound oligonucleotides (Tm; ΔG). However, these latter measures did not accurately predict the hierarchy of MSH ATPase activation. NMR studies of imino proton lifetime, solvent accessibility, and NOE connectivity suggest that sequence contexts that provoke improved MSH-activation displayed enhanced localized DNA flexibility: a dynamic DNA signature that may account for the wide range of lesions that activate MSH functions.