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Visualizing induced fit in early assembly of the human signal recognition particle

Nature Structural Biology volume 8, pages 515–520 (2001)Cite this article

Abstract

Assembly of almost all ribonucleoprotein complexes involves induced fit in the RNA and, thus, formation of one or more intermediate states. In assembly of the human signal recognition particle (SRP), we show that SRP19 binding to SRP RNA involves obligatory intermediates. An apparent discrepancy exists between the ratio of dissociation and association rate constants, determined in a partitioning experiment, and the equilibrium binding constant; this kinetic signature reflects formation of a stable intermediate in assembly of the ribonucleoprotein complex. Assembly intermediates were observed directly by time-resolved footprinting. SRP19 binds rapidly to SRP RNA to form an initial labile, but structurally specific, encounter complex involving both helices III and IV. Two subsequent steps of structural consolidation yield the native RNA–protein interface. SRP19 binding stabilizes helix IV in the region recognized by SRP54, consistent with protein–protein cooperativity mediated in part by mutual recognition of similar RNA structures. This mechanism illustrates principles general to ribonucleoprotein assembly reactions that rely on recruitment of architectural RNA binding proteins.

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Figure 1: Secondary structures of SRP RNA and the large subunit (LS) RNA.
Figure 2: Assembly, disassembly and equilibrium binding of the SRP19–RNA complex.
Figure 3: Visualization of SRP19–RNA complex formation by DMS and hydroxyl radical footprinting.
Figure 4: Time-resolved assembly of the SRP19–RNA complex.
Figure 5: Induced fit and cooperativity in early assembly of the human SRP.

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Acknowledgements

This work was supported by grants from the Searle Scholars Program of the Chicago Community Trust and by the NIH to K.M.W. We are indebted to K. Henry and H. Fried for gifts of plasmids and many helpful interactions in early phases of this work. We thank P. Bevilacqua and E. Westhof for helpful discussions; M. Been and T. Hall for careful readings of the manuscript; and L. Pedersen and L. Perera for assistance with RNA modeling.

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  1. Department of Chemistry, University of North Carolina, Chapel Hill, 27599-3290, North Carolina, USA

    Marsha A. Rose & Kevin M. Weeks

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  1. Marsha A. Rose
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Correspondence to Kevin M. Weeks.

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Rose, M., Weeks, K. Visualizing induced fit in early assembly of the human signal recognition particle. Nat Struct Mol Biol 8, 515–520 (2001). https://doi.org/10.1038/88577

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