title: Methylation of yeast ribosomal protein Rpl3 promotes translational elongation fidelity.
abstract: Rpl3, a highly conserved ribosomal protein, is methylated at histidine 243 by the Hpm1 methyltransferase in Saccharomyces cerevisiae. Histidine 243 lies close to the peptidyl transferase center in a functionally important region of Rpl3 designated as the basic thumb that coordinates the decoding, peptidyl transfer, and translocation steps of translation elongation. Hpm1 was recently implicated in ribosome biogenesis and translation. However, the biological role of methylation of its Rpl3 substrate has not been identified. Here we interrogate the role of Rpl3 methylation at H243 by investigating the functional impact of mutating this histidine residue to alanine (rpl3-H243A). Akin to Hpm1-deficient cells, rpl3-H243A cells accumulate 35S and 23S pre-rRNA precursors to a similar extent, confirming an important role for histidine methylation in pre-rRNA processing. In contrast, Hpm1-deficient cells but not rpl3-H243A mutants show perturbed levels of ribosomal subunits. We show that Hpm1 has multiple substrates in different subcellular fractions, suggesting that methylation of proteins other than Rpl3 may be important for controlling ribosomal subunit levels. Finally, translational fidelity assays demonstrate that like Hpm1-deficient cells, rpl3-H243A mutants have defects in translation elongation resulting in decreased translational accuracy. These data suggest that Rpl3 methylation at H243 is playing a significant role in translation elongation, likely via the basic thumb, but has little impact on ribosomal subunit levels. Hpm1 is therefore a multifunctional methyltransferase with independent roles in ribosome biogenesis and translation.
authors.affiliation: Department of Chemistry and Biochemistry and the Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Qais
authors.identifier: http://orcid.org/0000-0002-1715-9566
authors.initials: Q
authors.lastname: Al-Hadid
authors.affiliation: Department of Chemistry and Biochemistry and the Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Kevin
authors.identifier: http://orcid.org/0000-0001-8799-6207
authors.initials: K
authors.lastname: Roy
authors.affiliation: Department of Chemistry and Biochemistry and the Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Guillaume
authors.identifier: http://orcid.org/0000-0002-2505-1950
authors.initials: G
authors.lastname: Chanfreau
authors.affiliation: Department of Chemistry and Biochemistry and the Molecular Biology Institute, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: Steven G
authors.identifier: http://orcid.org/0000-0002-7303-6632
authors.initials: SG
authors.lastname: Clarke
mesh_terms: D006639:Histidine / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000502:physiology*; D010441:Peptide Chain Elongation, Translational; D011499:Protein Processing, Post-Translational; D012269:Ribosomal Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism* / Q000502:physiology*; D013379:Substrate Specificity
chemical_list: D012269:Ribosomal Proteins; C521718:Rpl3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; C000609573:Hpm1 protein, S cerevisiae; D008780:Methyltransferases
keywords: protein histidine methylation; ribosomal protein; ribosome biogenesis; translation elongation; translational fidelity
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