This is the main current project supported by an ERC Consolidator Grant 2015. We aim to understand the mechanism of action of SMC complexes, including understanding the role of SMC loaders and SMC accessory subunits, and how these proteins are regulated by ATP binding and hydrolysis for chromosome organisation.
We have done preliminary work in the characterisation of the SMC complex with AFM. Using an innovative approach that uses DNA as a fiducial marker to quantify volumes of proteins with high precision, we determined the oligomeric states and architecture of the Bacillus subtilis SMC complex. AFM results were used to color code the different protein components of the SMC complex: monomers of ScpA (pink); monomers and dimers of ScpB (green) ; ScpA-ScpB complexes (yellow); and SMC proteins (blue). The fiducial DNA molecule used in the study appears in white at the bottom part of the picture.
University of Bristol: Mark Dillingham Group
M.E. Fuentes-Perez et al.Biophysical Journal 102, 839-848 (2012).
Using DNA as a fiducial marker to study SMC complex interactions with the Atomic Force Micrsocope
M.E. Fuentes-Perez et al.METHODS 60, 113-121 (2013).
AFM volumetric methods for the characterization of proteins and nucleic acids
Taylor*, Pastrana* et al. Nucleic Acids Research 43(2), 719-731 (2015).
Specific and non-specific interactions of ParB with DNA: implications for chromosome segregation