3003 - Thomas Ve - Structural basis of Thoeris activation

Project ID:3003
RDM Collection:Q5241
ProjectType:Fee for Service
ProjectGroup:GRIFFITH - VE GROUP
Desc:Thoeris is a newly discovered bacterial NAD+ dependent antiphage defence system comprised of two components ThsB and ThsA. Phage infection triggers ThsB, which contains a TIR-domain, to produce an isomer of cyclic ADP-ribose (cADPR) which in turn activates ThsA. ThsA exhibits robust NAD+ cleavage and contains two domains, sirtuin-like and SLOG-like domains, and NAD depletion results in a growth arrest of the bacteria/cell death to prevent phage propagation. We have solved a low resolution (~3.5 A) ligand free ThsA crystal structure but co-crystallisation/soaking attempts have been unsuccessful. We are proposing to utilise cryoEM to resolve the complex of ThsA in complex with cADPR. We have already collected negatice staining images and mass photometry data showing a monodisperse tetramer in the presence of cADPR.

Aims/results:

We are proposing to utilise cryoEM to resolve the complex of ThsA in complex with cADPR. We have already collected negatice staining images and mass photometry data showing a monodisperse tetramer in the presence of cADPR.

AIM: [[sqote]]3 Angstrom resolution (or better) cryoEM structure of ThsA in complex with cADPR. Instruments: ARM 200 and 300[[sqote]]