Freitag vormittags Vorträge - Bunsentagung - Technische Universität ...

Freitag 21.05.2004, 10:45 Uhr
Plenarvortrag - Hauptthema
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Single Virus Tracing:
Visualization of the Infection Pathway of a Virus
into a Living Cell
Christoph Bräuchle
Department Chemie, Universität München, Butenandtstr. 11,
81377 München, Germany,
Christoph.Braeuchle@cup.uni-muenchen.de
Viruses play a major role in biology and medicine. A detailed
analysis of the different steps of a viral infection is not only
necessary to understand viral biology, but also for the development
of efficient antiviral drugs and safe viral gene therapy vectors.
Single Virus Tracing allows visualization of the infection
pathway of an individual virus particle labeled with fluorescent
dye molecules (even in the case of labeling with a single dye
molecule). The fluorescence of the marker molecule is imaged
and used to follow the pathway of the virus with high spatial
(≥ 40 nm) and time (≥ 10 ms) resolution. As a first model
system we have investigated the infection pathway of Adenoassociated
viruses (AAV) into HeLa cells. AAV shows promising
results for the use in gene therapy applications. A sequence
of events could be tracked, starting with the virus approaching
the cell surface, followed by receptor binding, membrane penetration,
endosome formation and trafficking, virus release from
the endosome and virus trafficking in the cytoplasm, as well as
penetration into the nuclear area. Besides AAV the studies have
been extended to other viruses like Human Immunodeficiency
Virus (HIV). HIV is an enveloped virus, which we have labeled
at its matrix protein (MA) and an internal accessory protein
(Vpr) by fusion with GFP or GFP variants, respectively. The investigations
focus on the interaction of HIV particles with the
plasma membrane of HeLa P4 cells and the subsequent membrane
fusion process.