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Projekt
Dissection of canonical and alternative pathways for human cytomegalovirus secondary envelopment using morphogenetic mutants and novel correlative microscopic methods
Antragsteller
Professor Dr. Thomas Mertens; Professor Dr. Paul Walther
Fachliche Zuordnung
Virologie
Förderung
Förderung von 2005 bis 2013
Projektkennung
Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 13122926
We want to combine existing specific expertises in electron microscopy and virology of human cytomegalovirus for better understanding of the envelopment mechanism of human cytomegalovirus by the new high-pressure freezing technique for electron microscopy. This new fixation method allows us to avoid artifacts of conventional chemical fixation. In preparatory work we already could show that thereby much more detailed pictures of cellular structures and of different viral particles can be obtained. The highly complex morphogenesis of this very large human ß-herpesvirus is still only partially understood and some aspects are controversially discussed. By generation of specific viral mutants with deleted or tagged tegument proteins, and of recombinant human cytomegaloviruses expressing homologous herpesvirus tegument genes, we want to clarify whether early processes of secondary tegumentation and envelopment represent a general mechanism of the herpesviridae family or are specific for herpesvirus subfamilies (a, ß, y). Furthermore using the UL83 (pp65 or lower matrix protein) deletion mutant we want to investigate, how viral particles, appearing normal in conventional EM, can be produced in the complete absence of the most abundant tegument protein (pp65) and how this protein, interspersed between envelope and inner tegument, is substituted to allow regular morphogenesis. We will use antibody labeling, electron tomography on thick plastic sections and cryo-scanning electron microscopy to clarify the involvement of cellular membrane structures, such as multivesicular bodies, in the process of tegumentation and envelopment.
DFG-Verfahren
Schwerpunktprogramme
