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Assays for the Interferon-Induced Enzyme 2′,5′ Oligoadenylate Synthetases

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Interferon Methods and Protocols

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 116))

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Abstract

Inhibition of protein synthesis by interferon treatment is mediated by two major pathways: the 2′–5′-linked oligoadenylates [2–5 (A)] synthetase-RNase L pathway and the double-stranded ribonucleic acid-dependent protein kinase-mediated pathway. 2–5 (A) synthetases are unique interferon-inducible enzymes that, upon activation by double-stranded RNA, polymerize adenosine triphosphate (ATP) to 2–5 (A) synthases. These 2–5 (A) synthetases bind and activate the latent RNase L, causing RNA degradation. In addition to the three major size classes of enzymatically active oligoadenylate synthetase proteins, at least one inactive oligoadenylate synthetase is known in human and mouse. Structure-function studies and recent crystal structure determination have identified several distinct sites in these proteins responsible for different biochemical functions. RNase L is the only known protein that binds to 2–5 (A) synthetases with very high affinity. Gene knockout studies of RNase L have identified its role in antiviral actions of interferon and in apoptosis. Recently, it has also been implicated in prostate cancer metastasis. In this chapter we describe several methodologies for studying biochemical and physiological properties of the 2–5 (A) synthetase-RNase L pathway.

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Acknowledgments

Relevant research in the authors’ laboratory was supported by the grant CA-68782 from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Molecular Biology, The Lerner Research Institute, Cleaveland Clinic Foundation, Cleveland, OH

    Saumendra N. Sarkar, Mitali Pandey & Ganes C. Sen

Authors
  1. Saumendra N. Sarkar
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  2. Mitali Pandey
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  3. Ganes C. Sen
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Editor information

Editors and Affiliations

  1. Department of Ophthalmology, The University of Oklahoma Health and Sciences Center, Oklahoma City, OK

    Daniel J. J. Carr

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© 2005 Humana Press Inc.

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Sarkar, S.N., Pandey, M., Sen, G.C. (2005). Assays for the Interferon-Induced Enzyme 2′,5′ Oligoadenylate Synthetases. In: Carr, D.J.J. (eds) Interferon Methods and Protocols. Methods in Molecular Medicine™, vol 116. Humana Press. https://doi.org/10.1385/1-59259-939-7:081

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  • DOI: https://doi.org/10.1385/1-59259-939-7:081

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-418-0

  • Online ISBN: 978-1-59259-939-4

  • eBook Packages: Springer Protocols

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