Abstract
During their life cycle viruses hijack host networks for their own needs. Discovering the basic biochemical interactions between viral proteins and host proteins is a crucial step towards understanding the viral life cycle. Due to the size of the host proteome high-throughput methods are needed to screen for such viral-host interactions. Despite significant efforts, conventional tools such as Yeast Two Hybrid largely failed. The failure is more pronounced when it comes to membrane associated proteins. We have designed PING, a sensitive high-throughput microfluidic assay that allows the screen of viral-host protein interactions as well as screen for inhibitors of such interactions. In a proof of principle experiment we screened the two HDV proteins and HCV NS5A against a hundred human proteins. We identified 7, 4 and 6 novel partners for sHDAg, LHDAg and NS5A, respectively. The average sensitivity of a single experiment (known partners identified out of all known partners included) was 62%, similar to bacterial interaction data previously measured with PING. The cumulative sensitivity in 4 experiments was 100%. These results pave the way for a whole proteome screen, which will allow us the horizontal overview we need into the biochemical network of viruses inside their host.
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call