Effect of murine cytomegalovirus on cellular protein expression for endosomal tubulation regulation

Citomegalovirus (CMV) u inficiranim stanicama uzrokuje opsežnu reorganizaciju sekretornih i endosomalnih membrana u svrhu stvaranja citoplazmatskog odjeljka za sklapanje viriona – cVAC (AC) (eng. cytoplasmic viral assembly compartment). Tijekom tog procesa, virus u stanici mijenja dinamiku i smještaj membranskih struktura djelujući na izražaj i aktivaciju regulatornih proteina koji se mobiliziraju na njihovu površinu. Rab10 GTPaza je jedan od mnogih takvih regulatornih proteina, a nalazimo ga na sučelju ranih endosoma, EE (eng. early endosomes) i endoplazmatskog reciklirajućeg odjeljka, ERC (eng. endoplasmic recycling compartment). U ovom diplomskom radu smo istražili učinak infekcije mišjim citomegalovirusom (MCMV) na izražaj i unutarstanični smještaj Rab10 i njegovih efektora, odnosno regulatora (Rab5, MICAL-L1, EHBP1, ACAP1, ACAP2, EHD1, EHD3) na inficiranim mišjim fibroblastima. Pokazali smo da se izražaj ispitivanih proteina ne mijenja tijekom infekcije, ali se Rab10 GTPaza i njegov efektor EHBP1 u kasnoj fazi infekcije nalaze nakupljeni u unutrašnjem dijelu AC (iAC). Prema tome, možemo zaključiti da MCMV infekcija uzrokuje reorganizaciju endosomalnih membrana na kojima se nalazi Rab10, ali ne mijenjajući njihov izražaj.Cytomegalovirus (CMV) causes extensive reorganization of the secretory and endosomal membranes in infected cells with a purpose of cytoplasmic viral assembly compartmens (cVAC/AC) formation. During that proces, CMV rearrange and change the dynamics of cellular membranes. One of the possible mechanisms involves targeting the expression of regulatory proteins that are activated and mobilized at surfaces of intracellular compartments. Rab10 GTPase represents one of many such proteins and can be found at the interface of early endosomes, EE and endoplasmic recycling compartment, ERC. In this thesis we have investigated the impact of infection with murine CMV (MCMV) on the Rab10 GTPase and its effectors and regulators (Rab5, MICAL-L1, EHBP1, ACAP1, ACAP2, EHD1, EHD3) expression and localization in the infected mouse fibroblasts. We have shown that their total expression do not change in the comparison with uninfected cells, but Rab10, and its effector, EHBP1, can be found in pericentriolar clusters inside AC (iAC) in the late phase of infection. Therefore, we can conclude that MCMV infection cause reorganization of Rab10 positive membranes, but without influence on their total expression