We are looking for a PhD
student (f/m) starting on 1 November 2022 or on 1 January 2023
in our team at the Institut de Biologie Structurale (IBS) in Grenoble, France working on the

Structure
of the vaccinia virus DNA replication machinery in complex with
DNA substrates by single-particle cryo-electron microscopy

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.

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.

The successful candidate will work in a small team interested in poxvirus DNA
replication directed by Prof. Wim
Burmeister within the « Viral Replication Machines group ». Cryo-EM and computing facilities are available
on the research campus, which is shared with international institutes: the EMBL Grenoble
outstation, the ESRF synchrotron and the ILL research
reactor. Working languages of the
institute are English and French. Grenoble is a mid-size
town with an international atmosphere located in the
middle of the French Alps. The net salary is about 1750 €/month (~2000 € gross salary),

In order to candidate please send a CV, a letter of
motivation and a copy of the Master diploma with marks to wim.burmeister@ibs.fr
before 30 September, 2022.

Bibliography

Tarbouriech N, Ducournau C,
Hutin S, Mas PJ, Man P, Forest E, Hart DJ, Peyrefitte CN,
Burmeister WP & Iseni F. The vaccinia virus DNA polymerase
structure provides insights into the mode of processivity factor
binding. Nat Commun. 2017;8: 1455.*
doi:10.1038/s41467-017-01542-z.

Bersch B, Tarbouriech N,
Burmeister WP, Iseni F. Solution structure of the C-terminal
domain of A20, the missing brick for the characterization of the
interface between vaccinia virus DNA polymerase and its
processivity factor. J Mol Biol. 2021; 167009.*
doi:10.1016/j.jmb.2021.167009.

Hutin, S., Ling, W.L.,
Tarbouriech, N., Schoehn, G, Grimm, G., Fischer, U. &
Burmeister, W.P. The vaccinia virus DNA helicase structure from
combined single-particle cryo-electron microscopy and AlphaFold2
prediction. Viruses. In press.

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