Thiouracil cross-linking mass spectrometry: a cell-based method to identify host factors involved in viral amplification.

ABSTRACT: Eukaryotic RNA viruses are known to utilize host factors; however, the identity of these factors and their role in the virus life cycle remain largely undefined. Here, we report a method to identify proteins bound to the viral RNA during amplification in cell culture: thiouracil cross-linking mass spectrometry (TUX-MS). TUX-MS relies on incorporation of a zero-distance cross-linker into the viral RNA during infection. Proteins bound to viral RNA are cross-linked prior to cell lysis, purified, and identified using mass spectrometry. Using the TUX-MS method, an unbiased screen for poliovirus (PV) host factors was conducted. All host and viral proteins that are known to interact with the poliovirus RNA were identified. In addition, TUX-MS identified an additional 66 host proteins that have no

SUBMITTER: Lenarcic EM 

PROVIDER: S-EPMC3719794 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature