Dokument

Zusammenfassung:
Das La Crosse-Virus (LACV) gehört zu der Familie der Bunyaviridae, eine der größten Virusfamilien, und wird unter anderem zusammen mit dem Prototyp Bunyamwera-Virus (BUNV) und dem Schmallenberg-Virus (SBV) in das Genus der Orthobunyaviren eingeteilt. Das Virus wird durch verschiedene Moskitospezies übertragen und stellt, aufgrund der Verteilung der Moskitos, im mittleren Westen und im Osten der USA ein Gesundheitsproblem dar. LACV kann eine schwere Enzephalitis und Meningitis auslösen, vor allem in Kindern und Jugendlichen unter 15 Jahren. Diese führen mitunter zu weitreichenden Folgen wie Lernschwierigkeiten, kognitive Defizite, Hyperaktivität, Krämpfe und Epilepsie. LACV gehört zu den Negativstrang-RNA-Viren mit einem tri-segmentiertem Genom und verfügt über einen Virulenzfaktor, das NSs (non-structural protein encoded on the S segment)-Protein, das die antivirale Interferon (IFN)-Induktion des infizierten Wirts inhibiert. Dies geschieht durch den gezielten proteasomalen Abbau der Rpb1-Untereinheit der RNA-Polymerase II (RNAPII). Ein solcher Abbau zeigt Ähnlichkeiten zu der DNA-Schadensantwort (DNA damage response, DDR), in der Rpb1 durch verschiedene Ubiquitin-E3-Ligasen ubiquitinyliert und somit dem Proteasom zugeführt wird. Hierbei spielt Nedd4 (neural precursor cell expressed developmentally down-regulated protein 4) eine tragende Rolle. In dieser Arbeit wurde gezeigt, dass das LACV-NSs-Protein eine generelle Ubiquitinylierung induziert, die Lysin48 (K48) verlinkt ist. Des Weiteren konnte gezeigt werden, dass im Speziellen die Rpb1-Untereinheit der RNAPII ubiquitinyliert wird. Zusätzlich wurde aufgezeigt, dass die Ubiquitin-E3-Ligase Nedd4 in den Abbau von Rpb1 involviert ist, da ein durch siRNA induzierter Nedd4-Knockdown den Abbau von Rpb1 und somit auch die Inhibition der IFN-Induktion teilweise aufhebt. Darüber hinaus konnte gezeigt werden, dass die subzelluläre Lokalisation von Nedd4 durch das LACV-NSs-Protein verändert wird, da Nedd4 in LACV-NSs-exprimierenden Zellen in perinukleären Strukturen akkumuliert, während es normalerweise homogen in der Zelle verteilt ist. Weiterhin wurde in dieser Arbeit die generelle Wirtszellantwort von LACV-infizierten Zellen mittels eines cDNA-Microarrays untersucht. Überraschenderweise stellte sich heraus, dass das LACV-NSs-Protein spezifisch Gene des angeborenen Immunsystems inhibiert, obwohl es bis dato nur als globaler Suppressor der RNAPII bekannt war.

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