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Analysis of SNARE-mediated Membrane Fusion Using an Enzymatic Cell Fusion Assay
Published on: October 19, 2012
Megaviruses contain various genes encoding for eukaryotic vesicle trafficking factors
Dany Khalifeh1, Emilie Neveu1, Dirk Fasshauer1
1Department of Computational Biology, University of Lausanne, Lausanne, Switzerland.
Abstract:
Many intracellular pathogens, such as bacteria and large viruses, enter eukaryotic cells via phagocytosis, then replicate and proliferate inside the host. To avoid degradation in the phagosomes, they have developed strategies to modify vesicle trafficking. Although several strategies of bacteria have been characterized, it is not clear whether viruses also interfere with the vesicle trafficking of the host. Recently, we came across SNARE proteins encoded in the genomes of several bacteria of the order Legionellales. These pathogenic bacteria may use SNAREs to interfere with vesicle trafficking, since SNARE proteins are the core machinery for vesicle fusion during transport. They assemble into membrane-bridging SNARE complexes that bring membranes together. We now have also discovered SNARE proteins in the genomes of diverse giant viruses. Our biochemical experiments showed that these proteins are able to form SNARE complexes. We also found other key trafficking factors that work together with SNAREs such as NSF, SM, and Rab proteins encoded in the genomes of giant viruses, suggesting that viruses can make use of a large genetic repertoire of trafficking factors. Most giant viruses possess different collections, suggesting that these factors entered the viral genome multiple times. In the future, the molecular role of these factors during viral infection need to be studied.
Insights
Giant viruses possess genes for SNARE proteins, crucial for vesicle trafficking. These viruses, like some bacteria, may manipulate host cell transport for their own replication and survival.
Area of Science:
- Microbiology
- Cell Biology
- Virology
Background:
- Intracellular pathogens, including bacteria and viruses, invade host cells via phagocytosis.
- Pathogens modify host vesicle trafficking to evade degradation within phagosomes.
- Bacterial strategies are known, but viral interference with vesicle trafficking remains unclear.
Purpose of the Study:
- To investigate whether giant viruses encode proteins involved in host vesicle trafficking.
- To explore the potential role of viral SNARE proteins in modulating cellular transport.
Main Methods:
- Bioinformatic analysis of giant virus genomes for trafficking-related genes.
- Biochemical experiments to assess the function of viral SNARE proteins.
- Identification of other viral trafficking factors like NSF, SM, and Rab proteins.
Main Results:
- Discovery of SNARE proteins and other key trafficking factors (NSF, SM, Rab) in diverse giant virus genomes.
- Biochemical validation that viral SNARE proteins can form functional SNARE complexes.
- Evidence suggests multiple independent acquisitions of these trafficking factors by viruses.
Conclusions:
- Giant viruses possess a genetic repertoire for manipulating host vesicle trafficking.
- Viral SNARE proteins are likely functional and contribute to the viral lifecycle.
- Further research is needed to elucidate the precise molecular mechanisms during infection.
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