Structural and functional analysis of the dna packaging protein ul89 from human cytomegalovirus
- Autores: Marta Nadal Rovira
- Directores de la Tesis: Maria Solà Vilarrubias (dir. tes.), Miquel Coll Capella (codir. tes.)
- Lectura: En la Universitat Autònoma de Barcelona ( España ) en 2010
- Idioma: español
- Tribunal Calificador de la Tesis: Ignacio Fita Rodríguez (presid.), Esteve Padrós i Morell (secret.), Darren Hart (voc.)
- Materias:
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- Resumen
Human cytomegalovirus (HCMV) is one of the eight human herpesvirus. Its infection rarely produces significant consequences in people with a competent immune system but can cause serious life threatening complications in immunocompromised people.
During viral replication, HCMV packages its DNA into the procapsid via the terminase complex that includes the UL89 protein. This complex nicks the newly-synthesized long DNA concatamers into genome units, suitable for encapsidation. Despite its high interest, no structural information is available for such herpesvirus encapsidation system.
This PhD dissertation covers the work done for the structural and functional analysis of the HCMV UL89 protein; from the gene cloning to the structure discussion.
Producing soluble and pure recombinant protein is a bottleneck for many interesting projects, as it was for UL89. With the goal of obtaining soluble full-length UL89, numerous strategies tried were unsuccessfully. However, using a new high throughput combinatory method, ESPRIT, we found a soluble domain of UL89. We could crystallize this construct and solve its three-dimensional structure at resolution of 2.15 Å. Finally, we performed activity assays to characterise its function.
