Structure of macromolecular assemblies

JLCarrascosa

 

José L. Carrascosa

Group Leader

Contact

 

Research summary

The activity of the group is focused in the study of macromolecular nanomachines that carry out defined biological functions. Among other systems, we are studying the molecular bases of assembly and maturation in viral systems.

 

Publications

Conesa JJ, Otón J, Chiappi M, Carazo JM, Pereiro E, Chichón FJ, Carrascosa JL. Intracellular nanoparticles mass quantification by near-edge absorption soft X-ray nanotomography. Sci Rep 2016; 6: 22354

Chiappi M, Conesa JJ, Pereiro E, Sorzano CO, Rodríguez MJ, Henzler K, Schneider G, Chichón FJ, Carrascosa JL. Cryo-soft X-ray tomography as a quantitative three-dimensional tool to model nanoparticle:cell interaction. J Nanobiotechnol 2016; 14: 15

Mertens J, Casado S, Mata CP, Hernando-Pérez M, de Pablo PJ, Carrascosa JL, Castón JR. A protein with simultaneous capsid scaffolding and dsRNA-binding activities enhances the birnavirus capsid mechanical stability. Sci Rep 2015; 5: 13486

González-García VA, Pulido-Cid M, Garcia-Doval C, Bocanegra R, van Raaij MJ, Martín-Benito J, Cuervo A, Carrascosa JL. Conformational Changes Leading to T7 DNA Delivery upon Interaction with the Bacterial Receptor. J Biol Chem 2015; 290: 10038-10044

Calero M, Chiappi M, Lazaro-Carrillo A, Rodríguez MJ, Chichón FJ, Crosbie-Staunton K, Prina-Mello A, Volkov Y, Villanueva A, Carrascosa JL. Characterization of interaction of magnetic nanoparticles with breast cancer cells. J Nanobiotechnol 2015; 13: 16


Funding

logo ministerio eco com2015-2017: Convocatoria 2014 Proyectos I+D. Programa Estatal de Fomento de la Investigación Científic y Técnica de Excelencia. BFU2014-54181-P: Correlación entre la estructura, ensamblaje y propiedades de las máquinas virales involucradas en procesos clave del ciclo viral.

2014-2017: Convocatoria Retos-Colaboración 2014. Programa Estatal de I+D+i Orientada a los Retos de la Sociedad. RTC-2014-1615-1: Desarrollo de una vacuna terapéutica frente al cáncer de cérvix basada en la proteína de fusión entre el dominio extra A de la fibronectina y el antígeno E7 del HPV.

COST logo 2 blue 72dpi

EU FLAG cmyk2013-2017:Acción COST MP 2017. EXTREMA: Enhanced X-ray Tomographic Reconstruction: Experiment, Modeling and Algorithms.

 

 

Fig1CarrascosaThe study of virus assembly allows us the molecular basis of macromolecular interactions with chemical and functional properties of the complexes assembled during the morphogenetic pathway of virus construction. Our group combines cryo-electron microscopy and computer three-dimensional image processing to obtain structures, at nanometric resolution, involved in bacteriophage T7 assembly. We solved two conformations of the DNA packaging motor (T7 large terminase), as well as several structures related to the DNA ejection complex (T7 tail), to obtain a detailed description of the DNA translocation process involved in the virus life cycle. Some components of these viruses (connector, terminase) are presently being used in synthetic environments to obtain delivery vehicles with improved specificity and efficiency.

Our structural studies are complemented with single-molecule analysis methods (atomic force microscopy, optical tweezers) to determine the mechanical properties of individual viral particles and subcomplexes at the nanoscopic scale. In this way, we associated the different mechanical behaviour of T7 intermediate maturation particles with their quasi-atomic structures. We also described macromolecular properties of various viral components and assemblies (dielectric constant of viral components, force development by molecular motors, differential properties of dsDNA and RNA).

We continued to extend the use of correlative microscopy by integration of electron tomographic methods with other microscopies, in particular with soft X-ray microscopy. The cryo-tomograms of whole, unperturbed cells infected with vaccinia virus obtained by X-ray microscopy showed previously unreported quantitative aspects of viral maturation and compartmentalisation. To exploit the possibilities of combined X-ray microscopy and analysis, we studied the interaction of magnetic nanoparticles and eukaryotic cells. Development of an approach that uses X-ray spectroscopy and tomography helped us to correlate the topological studies of cellular distribution of nanoparticles with the chemical quantitative information of metal uptake by cells.

 

Fig2Carrascosa

 grupoCararscosaRed

 

Name
Position
Contact
José L. Carrascosa Principal investigator
Rocío Arranz Senior Scientist
Ana Cuervo Gaspar Postdoctoral scientist
Rebeca Bocanegra Rojo Postdoctoral scientist
María José Rodríguez Gómez Postdoctoral scientist
Francisco Javier Chichón Postdoctoral scientist

 

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