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NRMT is an α-N-methyltransferase that methylates RCC1 and retinoblastoma protein

Nature volume 466, pages 1125–1128 (2010)Cite this article

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Abstract

The post-translational methylation of α-amino groups was first discovered over 30 years ago on the bacterial ribosomal proteins L16 and L33 (refs 1, 2), but almost nothing is known about the function or enzymology of this modification. Several other bacterial and eukaryotic proteins have since been shown to be α-N-methylated3,4,5,6,7,8,9,10. However, the Ran guanine nucleotide-exchange factor, RCC1, is the only protein for which any biological function of α-N-methylation has been identified3,11. Methylation-defective mutants of RCC1 have reduced affinity for DNA and cause mitotic defects3,11, but further characterization of this modification has been hindered by ignorance of the responsible methyltransferase. All fungal and animal N-terminally methylated proteins contain a unique N-terminal motif, Met-(Ala/Pro/Ser)-Pro-Lys, indicating that they may be targets of the same, unknown enzyme3,12. The initiating Met is cleaved, and the exposed α-amino group is mono-, di- or trimethylated. Here we report the discovery of the first α-N-methyltransferase, which we named N-terminal RCC1 methyltransferase (NRMT). Substrate docking and mutational analysis of RCC1 defined the NRMT recognition sequence and enabled the identification of numerous new methylation targets, including SET (also known as TAF-I or PHAPII) and the retinoblastoma protein, RB. Knockdown of NRMT recapitulates the multi-spindle phenotype seen with methylation-defective RCC1 mutants3, demonstrating the importance of α-N-methylation for normal bipolar spindle formation and chromosome segregation.

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Figure 1: Mettl11a is the α-N-RCC1 methyltransferase (NRMT).
Figure 2: Substrate recognition motif of NRMT.
Figure 3: NRMT methylates many targets, including SETα and RB.
Figure 4: Silencing NRMT reduces RCC1 association with chromatin, and increases frequency of mitotic defects.

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  • 26 August 2010

    A note was added in proof.

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Acknowledgements

This work was supported by research grants from the National Institutes of Health to I.G.M., D.F.H., and W.M.; C.E.S.T. was the recipient of a post-doctoral fellowship from the National Institutes of Health.

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Authors and Affiliations

  1. Department of Microbiology, Center for Cell Signaling, University of Virginia School of Medicine, Charlottesville, 22908, Virginia, USA

    Christine E. Schaner Tooley, Janusz J. Petkowski & Ian G. Macara

  2. Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, 22908, Virginia, USA

    Janusz J. Petkowski & Wladek Minor

  3. Department of Chemistry, University of Virginia, Charlottesville, 22904, Virginia, USA

    Tara L. Muratore-Schroeder, Jeremy L. Balsbaugh, Jeffrey Shabanowitz, Michal Sabat & Donald F. Hunt

  4. Department of Pathology, University of Virginia, Charlottesville, 22908, Virginia, USA

    Donald F. Hunt

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Contributions

C.E.S.T. purified NRMT and performed the experiments characterizing its function and targets and wrote the paper with I.G.M.; J.J.P. performed the ITC experiments and the RCC1 mutagenesis, and the docking experiments with the aid of M.S.; T.L.M.-S. and J.L.B. performed the mass spectrometry with J.S.; D.F.H. directed the mass spectrometry. W.M. coordinated the experiments of J.J.P.; I.G.M directed the biochemical and cell biological studies.

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Correspondence to Christine E. Schaner Tooley.

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The authors declare no competing financial interests.

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Schaner Tooley, C., Petkowski, J., Muratore-Schroeder, T. et al. NRMT is an α-N-methyltransferase that methylates RCC1 and retinoblastoma protein. Nature 466, 1125–1128 (2010). https://doi.org/10.1038/nature09343

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