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AKB: Biologische Physik

AKB 50: Imaging and Microscopy

AKB 50.9: Vortrag

Montag, 7. März 2005, 18:00–18:15, TU H2013

Single Virus Tracing: Real-time visualization of the membrane attachment and cellular uptake of individual HIV particles — •Thomas Endreß¹, Stefan Riegelsberger¹, Marko Lampe², Barbara Müller², Hans-Georg Käusslich², Don Lamb¹, and Christoph Bräuchle¹ — ¹Physikalische Chemie, Universität München, Butenandtstr. 11, 81377 München — ²Virologie, Unversität Heidelberg, Im Neuenheimer Feld 324, 69120 Heidelberg

Viruses play a major role in biology and medicine. A detailed analysis of the different steps of a viral infection is not only necessary for understanding viral biology, but also for the development of efficient antiviral drugs. Single Virus Tracing (SVT) allows visualization of the infection pathway of an individual virus labelled with fluorescent dye molecules. The fluorescence of the marker molecule is imaged and used to follow the pathway of the virus with high spatial (40 nm) and temporal (10 ms) resolution (Science, 294 (2001)1929).
HIV was labeled in the shell with its matrix protein (MA) using eGFP and in the core via its viral protein (Vpr) with mRFP. This allows simultaneous observation of the outer shell and subviral core of the individual HIV in real time. Analysis of virus-cell interactions by SVT revealed a detailed picture of the membrane interactions and cellular surface factors like HSPG involved in typical membrane attachment of HIVs. We were able to distinguish between the very rapid entry (≤ 1 min) of subviral particles by membrane fusion and a slower endosomal uptake of HIV (≥ 12 min). Furthermore, two types of intracellular trafficking were observed.