Research objectives and content
The overall aim of this project is to gain a thorough understanding of the molecular details of mismatch recognition by DNA repair enzymes. To do this, the interactions between E.coli MutS and synthetic DNA duplexes containing various mismatches and mutagenic bases will be estudied.The properties of the complete (96KDa) protein will be studied by a variety of biochemical techniques. In addition the structural details of MutS binding by preparing truncated derivatives (<30Kd) which retain mismatch binding activity will be examined. For this research programme, MutS protein and some regions of it will be synthesized and some techniques will be used to study the Interaction with mismatches in DNA: Footprinting, filter binding and gel retardation studies Ultraviolet melting (thermodynamic studies) Protein sequencing to determine the primary amino-acid sequence of MutS that binds to mismatches Crystallization of MutS and mini-proteins and their complexes with mismatches DNA
Training content (objective, benefit and expected impact)
This project will be a benefit for learning some of the techniques currently used in biochemistry assays. Having a better understanding of the features concerning the mismatches in DNA, could led to a closer knowledge of the causes and the therapy of cancer.
Links with industry / industrial relevance (22)