The Legionella pneumophila Methyltransferase RomA Methylates Also Non-histone Proteins during Infection
| Autor: | Srikanth Kudithipudi, Alexander Bröhm, Monica Rolando, Carmen Buchrieser, Sara Weirich, Albert Jeltsch, Maren Kirstin Schuhmacher |
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| Přispěvatelé: | Institute of Biochemistry and Technical Biochemistry = Institut für Biochemie und Technische Biochemie [Stuttgart], University of Stuttgart, Biologie des Bactéries intracellulaires - Biology of Intracellular Bacteria, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), This work has been supported by DFG (A.J., project JE252/7). Work in CB laboratory is financed by Institut Pasteur, the Centre national de la recherche scientifique, the Agency National de Recherche (ANR) (Grant No. ANR-10-LABEX-62-IBEID), the Infect-ERA project EUGENPATH (ANR-13-IFEC-0003-02) and the Fondation pour la Recherche Médicale (Grant No. DEQ20120323697)., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-13-IFEC-0003,EUGENPATH,Eukaryotic genes in vacuolar pathogens and symbionts - Implications for virulence, metabolism and ecology(2013), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Methyltransferase Legionella pneumophila protein lysine methyltransferase Substrate Specificity MESH: Legionella pneumophila / pathogenicity MESH: Legionnaires' Disease / metabolism Structural Biology Protein methylation Cloning Molecular MESH: Transcription Factors / chemistry Peptide sequence biology Chemistry Nuclear Proteins Methylation 3. Good health Histone MESH: HEK293 Cells MESH: Nuclear Proteins / chemistry Legionnaires' Disease MESH: Transcription Factors / metabolism MESH: Histone-Lysine N-Methyltransferase / metabolism MESH: Peptides / analysis non-histone substrate MESH: Peptides / genetics Protein domain MESH: Legionella pneumophila / enzymology MESH: Transcription Factors / genetics MESH: Methylation 03 medical and health sciences Histone H3 Bacterial Proteins Humans MESH: Cloning Molecular MESH: Nuclear Proteins / metabolism [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biochemistry [q-bio.BM] enzyme specificity protein methylation MESH: Bacterial Proteins / metabolism Molecular Biology MESH: Lysine / metabolism MESH: Humans 030102 biochemistry & molecular biology MESH: Nuclear Proteins / genetics Lysine Histone-Lysine N-Methyltransferase biology.organism_classification Molecular biology HEK293 Cells 030104 developmental biology MESH: Protein Processing Post-Translational MESH: HeLa Cells biology.protein MESH: Peptides / chemistry MESH: Substrate Specificity Peptides Protein Processing Post-Translational HeLa Cells Transcription Factors |
| Zdroj: | Journal of Molecular Biology Journal of Molecular Biology, 2018, 430 (13), pp.1912-1925. ⟨10.1016/j.jmb.2018.04.032⟩ Journal of Molecular Biology, Elsevier, 2018, 430 (13), pp.1912-1925. ⟨10.1016/j.jmb.2018.04.032⟩ |
| ISSN: | 0022-2836 1089-8638 |
| DOI: | 10.1016/j.jmb.2018.04.032⟩ |
| Popis: | International audience; RomA is a SET-domain containing protein lysine methyltransferase encoded by the Gram-negative bacterium Legionella pneumophila. It is exported into human host cells during infection and has been previously shown to methylate histone H3 at lysine 14 [Rolando et al. (2013), Cell Host Microbe, 13, 395-405]. Here, we investigated the substrate specificity of RomA on peptide arrays showing that it mainly recognizes a G-K-X-(PA) sequence embedded in a basic amino acid sequence context. Based on the specificity profile, we searched for possible additional RomA substrates in the human proteome and identified 34 novel peptide substrates. For nine of these, the corresponding full-length protein or protein domains could be cloned and purified. Using radioactive and antibody-based methylation assays, we showed that seven of them are methylated by RomA, four of them strongly, one moderately, and two weakly. Mutagenesis confirmed for the seven methylated proteins that methylation occurs at target lysine residues fitting to the specificity profile. Methylation of one novel substrate (AROS) was investigated in HEK293 cells overexpressing RomA and during infection with L. pneumophila. Methylation could be detected in both conditions, confirming that RomA methylates non-histone proteins in human cells. Our data show that the bacterial methyltransferase RomA methylates also human non-histone proteins suggesting a multifaceted role in the infection process. |
| Databáze: | OpenAIRE |
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