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The structure and oligomerization of the yeast arginine methyltransferase, Hmt1

Nature Structural Biology volume 7, pages 1165–1171 (2000)Cite this article

Abstract

Protein methylation at arginines is ubiquitous in eukaryotes and affects signal transduction, gene expression and protein sorting. Hmt1/Rmt1, the major arginine methyltransferase in yeast, catalyzes methylation of arginine residues in several mRNA-binding proteins and facilitates their export from the nucleus. We now report the crystal structure of Hmt1 at 2.9 Å resolution. Hmt1 forms a hexamer with approximate 32 symmetry. The surface of the oligomer is dominated by large acidic cavities at the dimer interfaces. Mutation of dimer contact sites eliminates activity of Hmt1 both in vivo and in vitro. Mutating residues in the acidic cavity significantly reduces binding and methylation of the substrate Npl3.

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Figure 1: Homology of arginine methyltransferases.
Figure 2: Hmt1-NΔ22+2 is functional.
Figure 3: Hmt1 monomer.
Figure 4: Ribbon representations of the Hmt1 dimer and hexamer.
Figure 5: Electrostatic surface potential of Hmt1 dimer, viewed from inside the hexamer (left, same view as Fig. 4a) and outside (right).
Figure 6: Replacement of the antenna region with eight alanines affects both in vivo function and migration of Hmt1 by gel filtration.
Figure 7: Crosslinking and gel filtration assays.
Figure 8: Differential activity and substrate-binding of Hmt1 truncation mutants.

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Acknowledgements

This work was supported by grants from NIH to P.A.S. and J.M.H., an NIH Post doctoral fellowship to A.E.M., an NSF Predoctoral fellowship to V.H.W and by the Giovanni Armenise-Harvard Center for Structural Biology. Diffraction data for this study were collected in collaboration with Michael Becker at Brookhaven National Laboratory in the Biology Department single crystal diffraction facility at beamline X12-C in the National Synchrotron Light Source. This facility is supported by the United States Department of Energy Offices of Health and Environmental Research and of Basic Energy Sciences, and by the National Institutes of Health National Center for Research Resources. We thank R. Sweet for synchrotron radiation facilities and assistance with data collection.

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Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Valerie H. Weiss, Anne E. McBride, Michelle A. Soriano, David J. Filman, Pamela A. Silver & James M. Hogle

  2. Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, 02115, Massachusetts, USA

    Anne E. McBride & Pamela A. Silver

  3. Committee on Higher Degrees in Biophysics, Harvard University, Cambridge, 02138 , Massachusetts, USA

    James M. Hogle

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  1. Valerie H. Weiss
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Correspondence to Pamela A. Silver or James M. Hogle.

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Weiss, V., McBride, A., Soriano, M. et al. The structure and oligomerization of the yeast arginine methyltransferase, Hmt1. Nat Struct Mol Biol 7, 1165–1171 (2000). https://doi.org/10.1038/82028

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