With the recent spread of monkeypox infections, poxviruses got into the headlines. Our team works on the elucidation of structure and function of the poxvirus DNA replication machinery, now mainly by single-particle cryo- electron microscopy. The DNA replication of vaccinia virus, a safe model system, involves the DNA polymerase holoenzyme complex E9-A20-D4, furthermore the hexameric helicase-primase D5. The aim is to determine structures of these assemblies by cryo-EM with bound DNA substrates, nucleotides, inhibitors and nucleotide analogues in order to stabilize different functional states for the understanding of poxvirus DNA replication. The project builds on the team's experiences in the structure determination and biophysical characterization of the partners and their domains, our recent results on the cryo-EM of the helicase domain of D5 and first results by cryo-EM on the holoenzyme.
Funding category: Contrat doctoral
Financé par l'ANR
PHD title: Doctorat de sciences du vivant
PHD Country: FranceOffer Requirements Specific Requirements
The candidate should be computer literate and have some experience in the use of Linux. Knowledge of relevant software packages used for cryo-EM will be an asset. Furthermore, a sound background in biochemistry, physics or biophysics is required as the project comprises multiple methods: protein production in the baculovirus-insect cell system; protein purification by affinity chromatography; sample preparation for cryo-EM; structure calculations for single-particle cryo-EM on a Linux computing cluster; analysis and visualization of protein structures.Contact Information