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| Identifier |
000370043 |
| Title |
Εντόπιση,οργάνωση και ρύθμιση του γονιδιώματος του κυτταρομεγαλοϊου ( CMV) κατά τη διάρκεια ενεργής λοίμωξης |
| Alternative Title |
Localization organization and dynamics of cytomegalovirus |
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Author
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Δημητροπούλου, Παναγιώτα Ν
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Thesis advisor
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Σπαντίδος,Δημήτριος
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Reviewer
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Σαμώνης, Γεώργιος
Σουρβίνος, Γεώργιος
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| Abstract |
Human Cytomegalovirus (HCMV) is a DNA herpes virus and a common human pathogen of considerable clinical interest. To image the IE1-72kDa HCMV protein a recombinant virus encoding IE1 fused to EGFP was constructed (CR401). Using this construct, the IE1-EGFP fusion was detected at ND10 within 2 h.p.i. and the complete disruption of ND10 imaged through to 6 h p.i. HCMV genomes and IE2-86K protein could be detected adjacent to the IE1-72K/ND10 foci. IE1-72K associates with metaphase chromatin, recruiting PML, Sp100 and STAT2. hDaxx, STAT1 and IE2-86K did not re-locate to metaphase chromatin; the fate of hDaxx is particularly important as this protein contributes to an intrinsic barrier to HCMV infection.
Since the progress of mitosis in infected cells has not thoroughly studied, we used the recombinant CR401 to visualize this dynamic phenomenon in living fibroblasts. Time-lapse microscopy in transiently expressing EGFP-IE1 and HcRed1-H2A HeLa cells revealed that IE1 was clearly associated with condensed chromatin at various stages of mitosis and eventually the IE1-expressing cells successfully completed cell division. Monitoring of individually infected fibroblasts by live cell imaging showed that a small proportion of cells initiated mitosis, progressing to prometaphase and metaphase quite readily. However, in the very few cells continuing to anaphase, lagging chromosomes were evident. A destruction of the mitotic spindle because of a HCMV expressed protein results in severe chromosome segregation defects.
We demonstrate here that the HCMV and EBV IE proteins differentially upregulate expression of ULBP2 and MICA/B, promoting NK cell activation. A panel of replication-deficient adenovirus recombinants encoding defined HCMV IE1 mutants has been generated, and is being used to characterize the mechanism responsible for promoting the ULBP2 response. The NKG2D receptor thus, appears to be exquisitely sensitive to herpesvirus IE gene expression.
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| Language |
Greek |
| Subject |
Bac technology |
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Condensed chromatin and mitotic block |
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Herpesviruses-Human cytomegalovirus |
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IE1-72KDA protein |
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IE2-86KDA protein |
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IE2-86KDA πρωτείνη |
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Immunology |
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Live cell microscopy |
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MICA/B |
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MICA/B |
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NKG2D ligands:ULBP1/2/3 |
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NKG2D πρόσδετες :ULBP1/2/3 |
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Nuclear domains ND10:PML-SP100-HDAXXproteins |
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Stat1-STAT2 proteins |
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Stat1-STAT2 πρωτείνες |
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Ερπητοίοι-ανθρώπινος κυτταρομεγαλοiος |
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Μικροσκοπία ζωντανών κυττάρων |
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Πυρηνικές δομές ND10:PML-SP100-HDAXX πρωτείνες |
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Συμπυκνωμένη χρωματίνη και διακοπή της μίτωσης |
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Τεχνολογία BAC |
| Issue date |
2010-07-20 |
| Collection
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School/Department--School of Medicine--Department of Medicine--Doctoral theses
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Type of Work--Doctoral theses
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| Views |
273 |
Εντόπιση,οργάνωση και ρύθμιση του γονιδιώματος του κυτταρομεγαλοϊου ( CMV) κατά τη διάρκεια ενεργής λοίμωξης
