Maintaining genomic integrity is important for any organism. DNA
mismatch repair (MMR) serves to correct errors that occur during DNA replication
and recombination, such as unpaired bases or mismatched bases. Mutl is a key
player and serves to coordinate protein-protein interactions. Recently it has been
shown that human Mutl functions as an endonuclease and that this activity is
imperative for functioning MMR. In this work, the X-ray crystal structure of the C-terminal
endonuclease domain of Bacillus subtilis Mutl (BsMutL-CTD) is
presented. Diffraction quality crystals of BsMutL-CTD were grown using vapor
diffusion. The crystal structure of BsMutL-CTD was solved using multiwavelength
anomalous diffraction. The structure reveals a putative metal binding
site which clusters closely in space with endonuclease motif. Using the structure
and sequence homology, several mutations were made and an investigation into
the endonuclease activity of BsMutL was performed. BsMutL was confirmed to
be a manganese-dependent endonuclease and key residues which contribute to
endonuclease function were identified. / Thesis / Master of Science (MSc)
| Identifer | oai:union.ndltd.org:mcmaster.ca/oai:macsphere.mcmaster.ca:11375/21627 |
| Date | 09 1900 |
| Creators | Lorenowicz, Jessica |
| Contributors | Guarne, Alba, Biochemistry |
| Source Sets | McMaster University |
| Language | English |
| Detected Language | English |
Page generated in 0.0129 seconds