AN EXAMINATION OF BINDING DIFFERENCES AMONG MISMATCH REPAIR RECOGNITION PROTEINS WITH MISMATCH AND CISPLATIN DAMAGED DNA

Mismatch repair is a major mechanism for the correction of replication errors in all organisms and increases the fidelity of DNA replication. Mismatch repair proteins have also demonstrated involvement in cellular response to the DNA damaging agent cisplatin. Cells deficient in mismatch repair have shown resistance to the cytotoxic effects of cisplatin treatment. Proposed mechanistic models for mismatch repair involvement require recognition of the damage site. To examine these models, E. coli MutS, S. cerevisae MSH2-MSH6, and MSH2-MSH3 were expressed to perform fluorescence anisotropy binding experiments and to crystallize the proteins in complex with cisplatin damaged DNA for comparison to mismatch DNA binding. All three mismatch repair (MMR) recognition complexes proved capable of binding cisplatin damaged DNA, but only MSH2-MSH6 demonstrated increased affinity towards cisplatin damage compared to undamaged DNA. Crystallization of a MMR protein with cisplatin damaged DNA was unsuccessful, but the methods presented here may be useful in further attempts.

Grantor dc:publisher

Wake Forest University

Year dc:date.issued

2010

Author dc:creator

• Rector, Brian

• DNA repair

Language dc:language.iso

en_US

Handle dc:identifier.uri

http://hdl.handle.net/10339/14920

OAI identifier oai:identifier

oai:wakespace.lib.wfu.edu:10339/14920