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The superhelical TPR-repeat domain of O-linked GlcNAc transferase exhibits structural similarities to importin α

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Abstract

Addition of N-acetylglucosamine (GlcNAc) is a ubiquitous form of intracellular glycosylation catalyzed by the conserved O-linked GlcNAc transferase (OGT). OGT contains an N-terminal domain of tetratricopeptide (TPR) repeats that mediates the recognition of a broad range of target proteins. Components of the nuclear pore complex are major OGT targets, as OGT depletion by RNA interference (RNAi) results in the loss of GlcNAc modification at the nuclear envelope. To gain insight into the mechanism of target recognition, we solved the crystal structure of the homodimeric TPR domain of human OGT, which contains 11.5 TPR repeats. The repeats form an elongated superhelix. The concave surface of the superhelix is lined by absolutely conserved asparagines, in a manner reminiscent of the peptide-binding site of importin α. Based on this structural similarity, we propose that OGT uses an analogous molecular mechanism to recognize its targets.

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Figure 1: Nucleoporins are major OGT targets.
Figure 2: Structure of the homodimeric TPR domain of human OGT.
Figure 3: The contiguous TPR repeats form a regular superhelical architecture.
Figure 4: The outer face of the TPR superhelix mediates homodimerization of OGT.
Figure 5: The inner surface of the TPR superhelix features a ladder of asparagines.
Figure 6: The inner surface of the TPR superhelix is similar to the peptide-binding surface of importin α.

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Acknowledgements

We are grateful to beamline scientists of X06SA at the Swiss Light Source (Zürich) and ESRF ID14-2 (Grenoble) for assistance during data collection. We thank M. Hothorn for introduction to data processing with XDS and P. Brick for critical reading of the manuscript. M.J. was supported by the Human Frontier Science Program Organization (RGP0063/2002-C).

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Authors and Affiliations

  1. Structural and Computational Biology, European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, D-69117, Germany

    Martin Jínek & Elena Conti

  2. Gene Expression Programmes, European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, D-69117, Germany

    Jan Rehwinkel & Elisa Izaurralde

  3. Laboratory of Cell Biochemistry and Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Brooke D Lazarus & John A Hanover

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  1. Martin Jínek
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  2. Jan Rehwinkel
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Correspondence to Elena Conti.

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Supplementary information

Supplementary Fig. 1

Nucleoporins are major OGT substrates in vivo. (PDF 734 kb)

Supplementary Methods (PDF 36 kb)

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Jínek, M., Rehwinkel, J., Lazarus, B. et al. The superhelical TPR-repeat domain of O-linked GlcNAc transferase exhibits structural similarities to importin α. Nat Struct Mol Biol 11, 1001–1007 (2004). https://doi.org/10.1038/nsmb833

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