Alfredo De Biasio
Molecular architecture and function of DNA replication and repair machines
The genetic information is encoded in long chains of deoxyribonucleic acid (DNA) molecules packaged into chromosomes in the cell nucleus. Before dividing, cells need to accurately duplicate their DNA, to ensure that each daughter cell has an identical copy. Critical players in DNA replication are the so-called DNA clamps, ring-shaped proteins that slide on the DNA double helix and anchor the polymerases, the enzymes that replicate DNA, to the genomic template. Beside DNA replication, sliding clamps function in the DNA damage response and the maintenance of genome integrity. Due to their central role in cell proliferation, DNA clamps are an important hallmark of tumours, as well as potential drug targets for anti-cancer therapy.
Our aim is to unravel the molecular architecture of eukaryotic DNA replication complexes involving DNA sliding clamps, and to shed light on their inner workings. To tackle these composite, supramolecular assemblies we integrate structural (X-ray, NMR, Electron Microscopy) and computational methods.
In our work, we have determined the structure of the human clamp PCNA bound to p15PAF, a modulator of the DNA damage tolerance pathway, revealing a unique mode of binding with functional implications. Our work has also provided a structural basis for the mechanism of PCNA sliding on DNA, resolving a long-lasting controversy on how eukaryotic clamps recognise and bind the DNA double helix.
An important focus of our current research is the integration of the structural studies with single-molecule studies, to resolve the spatio-temporal processes directed by the PCNA. To this purpose, we are collaborating with the laboratory of Jong-Bong Lee, at Postech University.
Lancey C, Tehseen M, Raducanu V-S, Rashid F, Merino N, Ragan TJ, Savva CG, Zaher MS, Shirbini A, Blanco FJ, Hamdan SM, De Biasio A. Structure of the processive human Pol δ holoenzyme. Nature Communications 11, 1109 (2020).
- De March, M. et al. (2017) 'Structural basis of human PCNA sliding on DNA.' Nature Communications, vol. 8. P. 13935.
- De Biasio, A et al. (2015) 'Structure of p15PAF‒PCNA complex and implications for clamp sliding during DNA replication and repair.' Nature Communications, vol. 6. P. 64392.