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Structure of free BglII reveals an unprecedented scissor-like motion for opening an endonuclease

Nature Structural Biology volume 8, pages 126–130 (2001)Cite this article

Abstract

Restriction endonuclease BglII completely encircles its target DNA, making contacts to both the major and minor grooves. To allow the DNA to enter and leave the binding cleft, the enzyme dimer has to rearrange. To understand how this occurs, we have solved the structure of the free enzyme at 2.3 Å resolution, as a complement to our earlier work on the BglII–DNA complex. Unexpectedly, the enzyme opens by a dramatic `scissor-like' motion, accompanied by a complete rearrangement of the α-helices at the dimer interface. Moreover, within each monomer, a set of residues — a ‘lever’ — lowers or raises to alternately sequester or expose the active site residues. Such an extreme difference in free versus complexed structures has not been reported for other restriction endonucleases. This elegant mechanism for capturing DNA may extend to other enzymes that encircle DNA.

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Figure 1: Overall structure.
Figure 2: Comparison of free and DNA bound monomers to show the lever motion.
Figure 3: A view of the scissor-like motion.

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Acknowledgements

We thank L. Berman and H. Lewis for facilitating data collection at NSLS. A.K.A. is supported by a grant from the NIH, and C.M.L. is supported by a Cancer Research Fund of the Damon Runyon-Walter Winchell Foundation Fellowship.

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Author notes

  1. Christine M. Lukacs, Rebecca Kucera & Ira Schildkraut

    Present address: New England Biolabs, 32 Tozer Road, Beverly, 01915, Massachusetts, USA

Authors and Affiliations

  1. Department of Physiology and Biophysics, Structural Biology Program, Mount Sinai School of Medicine, 1425 Madison Avenue, New York, 10029, New York, USA

    Christine M. Lukacs & Aneel K. Aggarwal

  2. Hoffman-La Roche Inc., 340 Kingsland St., Nutley, New Jersey, 07110, USA

    Christine M. Lukacs

Authors
  1. Christine M. Lukacs
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  2. Rebecca Kucera
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  4. Aneel K. Aggarwal
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Correspondence to Aneel K. Aggarwal.

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Lukacs, C., Kucera, R., Schildkraut, I. et al. Structure of free BglII reveals an unprecedented scissor-like motion for opening an endonuclease. Nat Struct Mol Biol 8, 126–130 (2001). https://doi.org/10.1038/84111

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