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Zawartość zarchiwizowana w dniu 2024-06-10

Modification of cell functions by human cytomegalovirus proteins in the absence of viral genome expression - role of incoming viral phosphoproteins 65 - ul83) and 71 - ul82


Research objectives and content
Infection with human Cytomegalovirus (HCMV) leads to profound, prolonged immuno-suppression which cannot be explained by lytic infection of immunocompetent cells. Results obtained in the host laboratory demonstrate that virion proteins can affect host cell metabolism and led to the hypothesis that incoming viral proteins may modify immune competent cell functions. The significance of incoming viral protein, pp65, both in vivo and in vitro, makes it important to define cellular proteins with which pp65 interacts in order to establish the role this protein plays in observed host-cell modifications. The transcriptional dependence of structural protein pp71 on pp65 synthesis requires investigating physical and biochemical interactions of these related phosphoproteins. We will develop expression vectors for both proteins with a tag epitope to allow (1) controlling the fate of tagged versus endogenous proteins produced during infection (2) performing co-precipitation experiments in uninfected cells transfected with recombinant expression vectors to determine normal cell proteins with which the viral proteins interact and (3) investigate biochemical modifications attributable to these proteins. Knowledge acquired and material developed should (1) help understand how HCMV modifies the functioning of immunocompetent cells in the absence of viral genome expression, (2) define functional interactions of viral pp65 and pp71with host-cell proteins and thus (3) might provide new targets for the intervention with antiviral drugs.
Training content (objective, benefit and expected impact)
Objective: to improve knowledge in virology and molecular biology of human cytomegalovirus (HCMV). Benefit: To specifically learn about the manipulation of eukariotic cell systems, expression and analysis of exogenous proteins and protein-protein physical and functional interactions. Expected impact: personally, to gain expertise in virology and molecular biology, to supplement current training in molecular biology of nucleic acids (Pavia, Italy) and in epidemiology and immunology (Stanford, USA); for the host laboratory, to contribute molecular biology expertise of HCMV necessary for the realization of their programs. Links with industry / industrial relevance (22): none

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