Determination of host RNA helicases activity in viral replication

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RNA helicases are encoded by all eukaryotic and prokaryotic cells and a minority of viruses. Activity of RNA helicases is necessary for all steps in the expression of cells and viruses and the host innate response to virus infection. Their vast functional repertoire is attributable to the core ATP-dependent helicase domain in conjunction with flanking domains that are interchangeable and engage viral and cellular cofactors. Here, we address the important issue of host RNA helicases that are necessary for replication of a virus. This chapter covers approaches to identification and characterization of candidate helicases and methods to define the biochemical and biophysical parameters of specificity and functional activity of the enzymes. We discuss the context of cellular RNA helicase activity and virion-associated RNA helicases. The methodology and choice of controls fosters the assessment of the virologic scope of RNA helicases across divergent cell lineages and viral replication cycles.

Original languageEnglish (US)
Pages (from-to)405-435
Number of pages31
JournalMethods in Enzymology
StatePublished - 2012

Bibliographical note

Funding Information:
We gratefully acknowledge Mr. Tim Vojt for illustrations, Dr. Amy Hayes, and Dr. Priya Kannian for helpful comments on this chapter, and Wei Jing for technical contributions in Fig. 19.1 . The authors appreciate the support of Pelotonia Predoctoral Fellowship, Comprehensive Cancer Center, Ohio State University to A. S. and NIH NCI P30CA100730 and RO1CA108882 to K. B. L.


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