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The DExH protein NPH-II is a processive and directional motor for unwinding RNA

Nature volume 403, pages 447–451 (2000)Cite this article

Abstract

All aspects of cellular RNA metabolism and processing involve DExH/D proteins, which are a family of enzymes that unwind or manipulate RNA in an ATP-dependent fashion1. DExH/D proteins are also essential for the replication of many viruses, and therefore provide targets for the development of therapeutics2. All DExH/D proteins characterized to date hydrolyse nucleoside triphosphates and, in most cases, this activity is stimulated by the addition of RNA or DNA1. Several members of the family unwind RNA duplexes in an NTP-dependent fashion in vitro1,3; therefore it has been proposed that DExH/D proteins couple NTP hydrolysis to RNA conformational change in complex macromolecular assemblies4. Despite the central role of DExH/D proteins, their mechanism of RNA helicase activity remains unknown. Here we show that the DExH protein NPH-II unwinds RNA duplexes in a processive, unidirectional fashion with a step size of roughly one-half helix turn. We show that there is a quantitative connection between ATP utilization and helicase processivity, thereby providing direct evidence that DExH/D proteins can function as molecular motors on RNA.

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Figure 1: Kinetics of duplex unwinding.
Figure 2: Estimation of the unwinding step size.
Figure 3: Probing helicase directionality with multipiece substrates (MPS).
Figure 4: Processivity of NPH-II.
Figure 5: The kinetic scheme for RNA helicase activity by NPH-II.

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Acknowledgements

We thank the members of the Pyle laboratory, particularly M. Boudvillain, J. B. Green and P. Z. Qin for many insightful discussions. E.J. was supported by a Curt-Engelhorn postdoctoral fellowship from the German Center for Cancer Research. A.M.P. is an assistant investigator of the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. The Department of Biochemistry and Molecular Biophysics, Columbia University, 630 W. 168th St, New York, 10032, New York, USA

    Eckhard Jankowsky & Anna Marie Pyle

  2. The Program in Molecular Biology, Sloan-Kettering Institute,

    Christian H. Gross & Stewart Shuman

  3. The Howard Hughes Medical Institute , 1275 York Ave., New York, 10021, New York , USA

    Anna Marie Pyle

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  1. Eckhard Jankowsky
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Jankowsky, E., Gross, C., Shuman, S. et al. The DExH protein NPH-II is a processive and directional motor for unwinding RNA. Nature 403, 447–451 (2000). https://doi.org/10.1038/35000239

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