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Approaching atomic resolution in structures of macromolecular assemblies by combining X-ray diffraction and electron microscopy : Methodological developments and application to SIRV1 virus

Objective

X-ray crystallography (XR) provides atomic resolution models of subunits of large biological assemblies, while electron microscopy (EM) allows lower-resolution imaging of whole macromolecular complexes. Combining the information from both techniques by fitting an atomic XR model into an EM density map enables in principle atomic-level interpretation of the EM data. We propose first to develop graphical software for manually fitting atomic XR models into EM data while simultaneously calculating the correlation between the models and the data using a fast reciprocal-space method [Navaza et al., Acta Crys. D58 (2002), 1820].

Second, we plan to implement an automatic fitting procedure with a complete 6-dimensional (3 rotations and 3 translations) search, based on a recently developed 6D extension of the fast rotational matching technique [Kovacs et al., Acta Crys. D59 (2003), 1371]. As a complement to these methodological developments, we will study using EM the structure of SIRV1, a rudivirus infecting thermophilic Archaea [Prangishvili et al., Genetics 152 (1999), 1387]. We will then use our newly developed software to fit the structures of viral proteins into the EM reconstruction.

Call for proposal

FP6-2004-MOBILITY-12
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Funding Scheme

EIF - Marie Curie actions-Intra-European Fellowships

Coordinator

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE
Address
3, Rue Michel Ange
Paris
France