Tags:
Biochemistry,
Molecular biology,
Molecular cell biology
Publications
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Schroer, Lisa; Weirich, Sara; Hammerstad, Marta; Hersleth, Hans-Petter; Grønsberg, Ida Andrietta & Hagen, Lars
[Show all 11 contributors for this article]
(2025).
Orthologues of the human protein histidine methyltransferase METTL9 display distinct substrate specificities.
Journal of Biological Chemistry.
ISSN 0021-9258.
301(7).
doi:
10.1016/j.jbc.2025.110318.
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The human (Homo sapiens; Hs) methyltransferase (MTase) METTL9 is the first enzyme shown to generate 1-methylhistidine (π-methylhistidine) in proteins. METTL9 preferentially methylates an alternating histidine (HxH) motif, where “x” is a small, uncharged amino acid, and multiple substrates have been identified. Putative METTL9 orthologues are found in most eukaryotes, and we have here investigated the activity of such enzymes from several species, representing all five eukaryotic supergroups. The majority of the tested enzymes demonstrated in vitro MTase activity on the prototype HsMETTL9 substrates ARMC6 and DNAJB12. We also detected protein methylation activity of the Caenorhabditis elegans METTL9 which had previously been suggested to be a DNA MTase. However, METTL9 from the fruit fly (Drosophila melanogaster; Dm) and the picoplankton Ostreococcus tauri (Ot) displayed distinct substrate specificities, differing from each other and from that of HsMETTL9. These differences were observed when recombinant proteins and short peptides were used as METTL9 substrates. To further analyze substrate specificity, we used peptide arrays to systematically replace the “x” residue and the residues flanking the HxH motif in a substrate peptide. This revealed varying degrees of tolerance among the METTL9 orthologues (Hs > Dm > Ot) for substitutions at these positions. Our results show that the METTL9 orthologues, although requiring an HxH target site, have evolved different substrate specificities, likely due to differing biologically relevant substrates in the respective organisms. Furthermore, we solved the X-ray structure of OtMETTL9, revealing several differences from the previously published HsMETTL9 structures that may explain its distinct substrate specificity.
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Malecki, Jedrzej Mieczyslaw; Weirich, Sara; Ramirez, Manuel; Hagen, Lars; Al-Egly, Jakin Saad & Anonsen, Jan Haug
[Show all 12 contributors for this article]
(2025).
Identification of substrates and sequence requirements for CARNMT1-mediated histidine methylation of C3H zinc fingers.
Journal of Biological Chemistry.
ISSN 0021-9258.
301(7).
doi:
10.1016/j.jbc.2025.110335.
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It has recently become clear that protein histidine methylation is widespread and functionally important in many cellular processes, and human CARNMT1 was recently reported as a novel protein histidine methyltransferase (HMT). We describe our independent uncovering of CARNMT1's protein HMT activity and a comprehensive assessment of its methylation targets and substrate specificity. Using a combination of in vitro methylation of cellular extracts and protein mass spectrometry, we identified several CARNMT1 substrates that were fully methylated in cells, all of which were C3H zinc finger (ZnF) proteins. These include the previously identified U2AF1, ZC3H15, and ZC3H18 but also the unreported RBM22, PPP1R10, PRR3, and RNF113A. Using peptide arrays, we investigated CARNMT1-mediated methylation of 145 candidate sequences, encompassing all C3H ZnFs and selected non-ZnFs. We found that only ∼30% of the tested sequences were methylated, with C3H ZnFs constituting the vast majority of the strongly methylated ones, most of which are also methylated in cells. This establishes peptide methylation as a good predictor of in vivo methylation. To investigate the specificity of CARNMT1, we systematically substituted His-proximal residues in four different substrate peptides. This generated four rather different sequence preference profiles, which were still quite restrictive for each peptide, indicating that substrate sequence recognition by CARNMT1 is context-dependent and that sequence-based prediction of additional CARNMT1 substrates may be challenging. We also identified several homologous methylation events in Caenorhabditis elegans and showed that they could be introduced by nematode CARNMT in vitro. Thus, CARNMT1 is an evolutionarily conserved protein HMT with a complex mode of substrate recognition.
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Willemen, Hanneke L.D.M.; Ribeiro, Patrícia Silva Santos; Broeks, Melissa; Meijer, Nils; Versteeg, Sabine & Tiggeler, Annefien
[Show all 11 contributors for this article]
(2023).
Inflammation-induced mitochondrial and metabolic disturbances in sensory neurons control the switch from acute to chronic pain.
Cell Reports Medicine.
4(11),
p. 1–24.
doi:
10.1016/j.xcrm.2023.101265.
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Pain often persists in patients with an inflammatory disease, even when inflammation has subsided. The molecular mechanisms leading to this failure in pain resolution and the transition to chronic pain are poorly understood. Mitochondrial dysfunction in sensory neurons links to chronic pain, but its role in resolution of inflammatory pain is unclear. Transient inflammation causes neuronal plasticity, called hyperalgesic priming, which impairs resolution of pain induced by a subsequent inflammatory stimulus. We identify that hyperalgesic priming in mice increases the expression of a mitochondrial protein (ATPSc-KMT) and causes mitochondrial and metabolic disturbances in sensory neurons. Inhibition of mitochondrial respiration, knockdown of ATPSCKMT expression, or supplementation of the affected metabolite is sufficient to restore resolution of inflammatory pain and prevents chronic pain development. Thus, inflammation-induced mitochondrial-dependent disturbances in sensory neurons predispose to a failure in resolution of inflammatory pain and development of chronic pain.
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Malecka, Agnieszka Maria; Østlie, Ingunn; Trøen, Gunhild; Malecki, Jedrzej Mieczyslaw; Delabie, Jan & Tierens, Anne
[Show all 9 contributors for this article]
(2023).
Gene expression analysis revealed downregulation of complement receptor 1 in clonal B cells in cold agglutinin disease.
Clinical and Experimental Immunology.
ISSN 0009-9104.
216(1),
p. 45–54.
doi:
10.1093/cei/uxad135.
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Abstract Cold agglutinin disease (CAD) is a rare B-cell lymphoproliferative disorder of the bone marrow, manifested by autoimmune hemolytic anemia caused by binding of monoclonal IgM autoantibodies to the I antigen. Underlying genetic changes have previously been reported, but their impact on gene expression profile has been unknown. Here, we define differentially expressed genes in CAD B cells. To unravel downstream alteration in cellular pathways, gene expression by RNA sequencing was undertaken. Clonal B-cell samples from 12 CAD patients and IgM-expressing memory B cells from 4 healthy individuals were analyzed. Differential expression analysis and filtering resulted in 93 genes with significant differential expression. Top upregulated genes included SLC4A1, SPTA1, YBX3, TESC, HBD, AHSP, TRAF1, HBA2, RHAG, CA1, SPTB, IL10, UBASH3B, ALAS2, HBA1, CRYM, RGCC, KANK2, and IGHV4-34. They were upregulated at least 8-fold, while complement receptor 1 (CR1/CD35) was downregulated 11-fold in clonal CAD B cells compared to control B cells. Flow cytometry analyses further confirmed reduced CR1 (CD35) protein expression by clonal CAD IgM+ B cells compared to IgM+ memory B cells in controls. CR1 (CD35) is an important negative regulator of B-cell activation and differentiation. Therefore, reduced CR1 (CD35) expression may increase activation, proliferation, and antibody production in CAD-associated clonal B cells.
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Falnes, Pål; Malecki, Jedrzej Mieczyslaw; Gonzalez-Molina, Maria del Carmen Herrera; Bengtsen, Mads & Davydova, Erna
(2023).
Human seven-β-strand (METTL) methyltransferases - conquering the universe of protein lysine methylation.
Journal of Biological Chemistry.
ISSN 0021-9258.
299(6),
p. 1–15.
doi:
10.1016/j.jbc.2023.104661.
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Lysine methylation is an abundant posttranslational modification, which has been most intensively studied in the context of histone proteins, where it represents an important epigenetic mark. Lysine methylation of histone proteins is primarily catalyzed by SET-domain methyltransferases (MTases). However, it has recently become evident that also another MTase family, the so-called seven-β-strand (7BS) MTases, often denoted METTLs (methyltransferase-like), contains several lysine (K)-specific MTases (KMTs). These enzymes catalyze the attachment of up to three methyl groups to lysine residues in specific substrate proteins, using S-adenosylmethionine (AdoMet) as methyl donor. About a decade ago, only a single human 7BS KMT was known, namely the histone-specific DOT1L, but 15 additional 7BS KMTs have now been discovered and characterized. These KMTs typically target a single nonhistone substrate that, in most cases, belongs to one of the following three protein groups: components of the cellular protein synthesis machinery, mitochondrial proteins, and molecular chaperones. This article provides an extensive overview and discussion of the human 7BS KMTs and their biochemical and biological roles.
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Engelfriet, Melanie Lianne; Malecki, Jedrzej Mieczyslaw; Forsberg, Frida; Falnes, Pål & Ciosk, Rafal
(2023).
Characterization of the biochemical activity and tumor-promoting role of the dual protein methyltransferase METL-13/METTL13 in Caenorhabditis elegans.
PLOS ONE.
18(6),
p. 1–16.
doi:
10.1371/journal.pone.0287558.
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The methyltransferase-like protein 13 (METTL13) methylates the eukaryotic elongation factor 1 alpha (eEF1A) on two locations: the N-terminal amino group and lysine 55. The absence of this methylation leads to reduced protein synthesis and cell proliferation in human cancer cells. Previous studies showed that METTL13 is dispensable in non-transformed cells, making it potentially interesting for cancer therapy. However, METTL13 has not been examined yet in whole animals. Here, we used the nematode Caenorhabditis elegans as a simple model to assess the functions of METTL13. Using methyltransferase assays and mass spectrometry, we show that the C . elegans METTL13 (METL-13) methylates eEF1A (EEF-1A) in the same way as the human protein. Crucially, the cancer-promoting role of METL-13 is also conserved and depends on the methylation of EEF-1A, like in human cells. At the same time, METL-13 appears dispensable for animal growth, development, and stress responses. This makes C . elegans a convenient whole-animal model for studying METL13-dependent carcinogenesis without the complications of interfering with essential wild-type functions.
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Malecki, Jedrzej Mieczyslaw; Davydova, Erna & Falnes, Pål
(2022).
Protein methylation in mitochondria.
Journal of Biological Chemistry.
ISSN 0021-9258.
298(4).
doi:
10.1016/j.jbc.2022.101791.
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Many proteins are modified by posttranslational methylation, introduced by a number of methyltransferases (MTases). Protein methylation plays important roles in modulating protein function and thus in optimizing and regulating cellular and physiological processes. Research has mainly focused on nuclear and cytosolic protein methylation, but it has been known for many years that also mitochondrial proteins are methylated. During the last decade, significant progress has been made on identifying the MTases responsible for mitochondrial protein methylation and addressing its functional significance. In particular, several novel human MTases have been uncovered that methylate lysine, arginine, histidine, and glutamine residues in various mitochondrial substrates. Several of these substrates are key components of the bioenergetics machinery, e.g., respiratory Complex I, citrate synthase, and the ATP synthase. In the present review, we report the status of the field of mitochondrial protein methylation, with a particular emphasis on recently discovered human MTases. We also discuss evolutionary aspects and functional significance of mitochondrial protein methylation and present an outlook for this emergent research field.
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Pekec, Tina; Lewandowski, Jarosław; Komur, Alicja; Sobańska, Daria; Guo, Yanwu & Świtońska-Kurkowska, Karolina
[Show all 12 contributors for this article]
(2022).
Ferritin-mediated iron detoxification promotes hypothermia survival in Caenorhabditis elegans and murine neurons.
Nature Communications.
13(1).
doi:
10.1038/s41467-022-32500-z.
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Abstract How animals rewire cellular programs to survive cold is a fascinating problem with potential biomedical implications, ranging from emergency medicine to space travel. Studying a hibernation-like response in the free-living nematode Caenorhabditis elegans , we uncovered a regulatory axis that enhances the natural resistance of nematodes to severe cold. This axis involves conserved transcription factors, DAF-16/FoxO and PQM-1, which jointly promote cold survival by upregulating FTN-1, a protein related to mammalian ferritin heavy chain (FTH1). Moreover, we show that inducing expression of FTH1 also promotes cold survival of mammalian neurons, a cell type particularly sensitive to deterioration in hypothermia. Our findings in both animals and cells suggest that FTN-1/FTH1 facilitates cold survival by detoxifying ROS-generating iron species. We finally show that mimicking the effects of FTN-1/FTH1 with drugs protects neurons from cold-induced degeneration, opening a potential avenue to improved treatments of hypothermia.
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Malecki, Jedrzej Mieczyslaw; O'Donohue, Marie-Francoise; Kim, Yeji; Jakobsson, Magnus; Gessa, Luca & Pinto, Rita
[Show all 14 contributors for this article]
(2021).
Human METTL18 is a histidine-specific methyltransferase that targets RPL3 and affects ribosome biogenesis and function.
Nucleic Acids Research (NAR).
ISSN 0305-1048.
49(6),
p. 3185–3203.
doi:
10.1093/nar/gkab088.
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Abstract Protein methylation occurs primarily on lysine and arginine, but also on some other residues, such as histidine. METTL18 is the last uncharacterized member of a group of human methyltransferases (MTases) that mainly exert lysine methylation, and here we set out to elucidate its function. We found METTL18 to be a nuclear protein that contains a functional nuclear localization signal and accumulates in nucleoli. Recombinant METTL18 methylated a single protein in nuclear extracts and in isolated ribosomes from METTL18 knockout (KO) cells, identified as 60S ribosomal protein L3 (RPL3). We also performed an RPL3 interactomics screen and identified METTL18 as the most significantly enriched MTase. We found that His-245 in RPL3 carries a 3-methylhistidine (3MH; τ-methylhistidine) modification, which was absent in METTL18 KO cells. In addition, both recombinant and endogenous METTL18 were found to be automethylated at His-154, thus further corroborating METTL18 as a histidine-specific MTase. Finally, METTL18 KO cells displayed altered pre-rRNA processing, decreased polysome formation and codon-specific changes in mRNA translation, indicating that METTL18-mediated methylation of RPL3 is important for optimal ribosome biogenesis and function. In conclusion, we have here established METTL18 as the second human histidine-specific protein MTase, and demonstrated its functional relevance.
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Davydova, Erna; Shimazu, Tadahiro; Schuhmacher, Maren Kirstin; Jakobsson, Magnus E.; Willemen, Hanneke L.D.M. & Liu, Tongri
[Show all 28 contributors for this article]
(2021).
The methyltransferase METTL9 mediates pervasive 1-methylhistidine modification in mammalian proteomes.
Nature Communications.
12,
p. 1–14.
doi:
10.1038/s41467-020-20670-7.
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Abstract Post-translational methylation plays a crucial role in regulating and optimizing protein function. Protein histidine methylation, occurring as the two isomers 1- and 3-methylhistidine (1MH and 3MH), was first reported five decades ago, but remains largely unexplored. Here we report that METTL9 is a broad-specificity methyltransferase that mediates the formation of the majority of 1MH present in mouse and human proteomes. METTL9-catalyzed methylation requires a His-x-His (HxH) motif, where “x” is preferably a small amino acid, allowing METTL9 to methylate a number of HxH-containing proteins, including the immunomodulatory protein S100A9 and the NDUFB3 subunit of mitochondrial respiratory Complex I. Notably, METTL9-mediated methylation enhances respiration via Complex I, and the presence of 1MH in an HxH-containing peptide reduced its zinc binding affinity. Our results establish METTL9-mediated 1MH as a pervasive protein modification, thus setting the stage for further functional studies on protein histidine methylation.
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Pinto, Rita; Vågbø, Cathrine Broberg; Jakobsson, Magnus; Yeji, Kim; Baltissen, Marijke P & O'Donohue, Marie-Francoise
[Show all 16 contributors for this article]
(2019).
The human methyltransferase ZCCHC4 catalyses N6-methyladenosine modification of 28S ribosomal RNA.
Nucleic Acids Research (NAR).
ISSN 0305-1048.
48(2),
p. 830–846.
doi:
10.1093/nar/gkz1147.
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RNA methylations are essential both for RNA structure and function, and are introduced by a number of distinct methyltransferases (MTases). In recent years, N6-methyladenosine (m6A) modification of eukaryotic mRNA has been subject to intense studies, and it has been demonstrated that m6A is a reversible modification that regulates several aspects of mRNA function. However, m6A is also found in other RNAs, such as mammalian 18S and 28S ribosomal RNAs (rRNAs), but the responsible MTases have remained elusive. 28S rRNA carries a single m6A modification, found at position A4220 (alternatively referred to as A4190) within a stem–loop structure, and here we show that the MTase ZCCHC4 is the enzyme responsible for introducing this modification. Accordingly, we found that ZCCHC4 localises to nucleoli, the site of ribosome assembly, and that proteins involved in RNA metabolism are overrepresented in the ZCCHC4 interactome. Interestingly, the absence of m6A4220 perturbs codon-specific translation dynamics and shifts gene expression at the translational level. In summary, we establish ZCCHC4 as the enzyme responsible for m6A modification of human 28S rRNA, and demonstrate its functional significance in mRNA translation.
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Malecki, Jedrzej Mieczyslaw; Willemen, Hanneke L.D.M.; Pinto, Rita; Ho, Angela Yeuan Yen; Moen, Anders & Kjønstad, Ingrid Fadum
[Show all 10 contributors for this article]
(2018).
Lysine methylation by the mitochondrial methyltransferase FAM173B optimizes the function of mitochondrial ATP synthase.
Journal of Biological Chemistry.
ISSN 0021-9258.
294(4),
p. 1128–1141.
doi:
10.1074/jbc.RA118.005473.
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Lysine methylation is an important post-translational modification that is also present on mitochondrial proteins, but the mitochondrial lysine-specific methyltransferases (KMTs) responsible for modification are in most cases unknown. Here, we set out to determine the function of human family with sequence similarity 173 member B (FAM173B), a mitochondrial methyltransferase (MTase) reported to promote chronic pain. Using bioinformatics analyses and biochemical assays, we found that FAM173B contains an atypical, noncleavable mitochondrial targeting sequence responsible for its localization to mitochondria. Interestingly, CRISPR/Cas9-mediated KO of FAM173B in mammalian cells abrogated trimethylation of Lys-43 in ATP synthase c-subunit (ATPSc), a modification previously reported as ubiquitous among metazoans. ATPSc methylation was restored by complementing the KO cells with enzymatically active human FAM173B or with a putative FAM173B orthologue from the nematode Caenorhabditis elegans. Interestingly, lack of Lys-43 methylation caused aberrant incorporation of ATPSc into the ATP synthase complex and resulted in decreased ATP-generating ability of the complex, as well as decreased mitochondrial respiration. In summary, we have identified FAM173B as the long-sought KMT responsible for methylation of ATPSc, a key protein in cellular ATP production, and have demonstrated functional significance of ATPSc methylation. We suggest renaming FAM173B to ATPSc-KMT (gene name ATPSCKMT).
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Jakobsson, Magnus; Malecki, Jedrzej Mieczyslaw; Halabelian, Levon; Nilges, Benedikt S.; Pinto, Rita & Kudithipudi, Srikanth
[Show all 14 contributors for this article]
(2018).
The dual methyltransferase METTL13 targets N terminus and Lys55 of eEF1A and modulates codon-specific translation rates.
Nature Communications.
9:3411,
p. 1–15.
doi:
10.1038/s41467-018-05646-y.
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Eukaryotic elongation factor 1 alpha (eEF1A) delivers aminoacyl-tRNA to the ribosome and thereby plays a key role in protein synthesis. Human eEF1A is subject to extensive post-translational methylation, but several of the responsible enzymes remain unknown. Using a wide range of experimental approaches, we here show that human methyltransferase (MTase)-like protein 13 (METTL13) contains two distinct MTase domains targeting the N terminus and Lys55 of eEF1A, respectively. Our biochemical and structural analyses provide detailed mechanistic insights into recognition of the eEF1A N terminus by METTL13. Moreover, through ribosome profiling, we demonstrate that loss of METTL13 function alters translation dynamics and results in changed translation rates of specific codons. In summary, we here unravel the function of a human MTase, showing that it methylates eEF1A and modulates mRNA translation in a codon-specific manner.
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Malecki, Jedrzej Mieczyslaw; Jakobsson, Magnus; Ho, Angela Yeuan Yen; Moen, Anders; Rustan, Arild & Falnes, Pål
(2017).
Uncovering human METTL12 as a mitochondrial methyltransferase that modulates citrate synthase activity through metabolite-sensitive lysine methylation.
Journal of Biological Chemistry.
ISSN 0021-9258.
292(43),
p. 17950–17962.
doi:
10.1074/jbc.M117.808451.
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Lysine methylation is an important and much-studied posttranslational modification of nuclear and cytosolic proteins but is present also in mitochondria. However, the responsible mitochondrial lysine-specific methyltransferases (KMTs) remain largely elusive. Here, we investigated METTL12, a mitochondrial human S-adenosylmethionine (AdoMet)-dependent methyltransferase and found it to methylate a single protein in mitochondrial extracts, identified as citrate synthase (CS). Using several in vitro and in vivo approaches, we demonstrated that METTL12 methylates CS on Lys-395, which is localized in the CS active site. Interestingly, the METTL12-mediated methylation inhibited CS activity and was blocked by the CS substrate oxaloacetate. Moreover, METTL12 was strongly inhibited by the reaction product S-adenosylhomocysteine (AdoHcy). In summary, we have uncovered a novel human mitochondrial KMT that introduces a methyl modification into a metabolic enzyme and whose activity can be modulated by metabolic cues. Based on the established naming nomenclature for similar enzymes, we suggest that METTL12 be renamed CS-KMT (gene name CSKMT). This research was originally published in the Journal of Biological Chemistry. © 2017 American Society for Biochemistry and Molecular Biology
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Malecki, Jedrzej Mieczyslaw; Aileni, Vinay Kumar; Ho, Angela Yeuan Yen; Schwarz, Juliane; Moen, Anders & Sørensen, Vigdis
[Show all 10 contributors for this article]
(2017).
The novel lysine specific methyltransferase METTL21B affects mRNA translation through inducible and dynamic methylation of Lys-165 in human eukaryotic elongation factor 1 alpha (eEF1A).
Nucleic Acids Research (NAR).
ISSN 0305-1048.
45(8),
p. 4370–4389.
doi:
10.1093/nar/gkx002.
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Lysine methylation is abundant on histone proteins, representing a dynamic regulator of chromatin state and gene activity, but is also frequent on many nonhistone proteins, including eukaryotic elongation factor 1 alpha (eEF1A). However, the functional significance of eEF1Amethylation remains obscure and it has remained unclear whether eEF1A methylation is dynamic and subject to active regulation. We here demonstrate, using a wide range of in vitro and in vivo approaches, that the previously uncharacterized human methyltransferase METTL21B specifically targets Lys-165 in eEF1A in an aminoacyl-tRNA- and GTP-dependent manner. Interestingly, METTL21B mediated eEF1A methylation showed strong variation across different tissues and cell lines, and was induced by altering growth conditions or by treatment with certain ER-stress-inducing drugs, concomitant with an increase in METTL21B gene expression. Moreover, genetic ablation of METTL21B function in mammalian cells caused substantial alterations in mRNA translation, as measured by ribosomal profiling. A non-canonical function for eEF1A in organization of the cellular cytoskeleton has been reported, and interestingly, METTL21B accumulated in centrosomes, in addition to the expected cytosolic localization. In summary, the present study identifies METTL21B as the enzyme responsible for methylation of eEF1A on Lys-165 and shows that this modification is dynamic, inducible and likely of regulatory importance.
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Jakobsson, Magnus; Malecki, Jedrzej Mieczyslaw; Nilges, Benedikt S.; Moen, Anders; Leidel, Sebastian A. & Falnes, Pål
(2017).
Methylation of human eukaryotic elongation factor
alpha (eEF1A) by a member of a novel protein lysine
methyltransferase family modulates mRNA translation.
Nucleic Acids Research (NAR).
ISSN 0305-1048.
45,
p. 8239–8254.
doi:
10.1093/nar/gkx432.
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Many cellular proteins are methylated on lysine residues and this has been most intensively studied for histone proteins. Lysine methylations on non-histone proteins are also frequent, but in most cases the functional significance of the methylation event, as well as the identity of the responsible lysine (K) specific methyltransferase (KMT), remain unknown. Several recently discovered KMTs belong to the so-called seven-β-strand (7BS) class of MTases and we have here investigated an uncharacterized human 7BS MTase currently annotated as part of the endothelin converting enzyme 2, but which should be considered a separate enzyme. Combining in vitro enzymology and analyzes of knockout cells, we demonstrate that this MTase efficiently methylates K36 in eukaryotic translation elongation factor 1 alpha (eEF1A) in vitro and in vivo. We suggest that this novel KMT is named eEF1A-KMT4 (gene name EEF1AKMT4), in agreement with the recently established nomenclature. Furthermore, by ribosome profiling we show that the absence of K36 methylation affects translation dynamics and changes translation speed of distinct codons. Finally, we show that eEF1A-KMT4 is part of a novel family of human KMTs, defined by a shared sequence motif in the active site and we demonstrate the importance of this motif for catalytic activity.
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Falnes, Pål; Jakobsson, Magnus; Davydova, Erna; Ho, Angela Yeuan Yen & Malecki, Jedrzej Mieczyslaw
(2016).
Protein lysine methylation by seven-β-strand methyltransferases.
Biochemical Journal.
ISSN 0264-6021.
473(14),
p. 1995–2009.
doi:
10.1042/BCJ20160117.
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Malecki, Jedrzej Mieczyslaw; Dahl, Helge-Andre; Moen, Anders; Davydova, Erna & Falnes, Pål
(2016).
The METTL20 homologue from agrobacterium tumefaciens is a dual specificity protein-lysine methyltransferase that targets ribosomal protein L7/L12 and the β subunit of electron transfer flavoprotein (ETFβ).
Journal of Biological Chemistry.
ISSN 0021-9258.
291(18),
p. 9581–9595.
doi:
10.1074/jbc.M115.709261.
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Jakobsson, Magnus; Davydova, Erna; Malecki, Jedrzej Mieczyslaw; Moen, Anders & Falnes, Pål
(2015).
Saccharomyces cerevisiae eukaryotic elongation factor 1A (eEF1A) is methylated at Lys-390 by a METTL21-like methyltransferase.
PLOS ONE.
10(6).
doi:
10.1371/journal.pone.0131426.
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The human methyltransferases (MTases) METTL21A and VCP-KMT (METTL21D) were recently shown to methylate single lysine residues in Hsp70 proteins and in VCP, respectively. The yet uncharacterized MTase encoded by the YNL024C gene in Saccharomyces cerevisiae shows high sequence similarity to METTL21A and VCP-KMT, as well as to their uncharacterized paralogues METTL21B and METTL21C. Despite being most similar to METTL21A, the Ynl024c protein does not methylate yeast Hsp70 proteins, which were found to be unmethylated on the relevant lysine residue. Eukaryotic translation elongation factor eEF1A in yeast has been reported to contain four methylated lysine residues (Lys30, Lys79, Lys318 and Lys390), and we here show that the YNL024C gene is required for methylation of eEF1A at Lys390, the only of these methylations for which the responsible MTase has not yet been identified. Lys390 was found in a partially monomethylated state in wild-type yeast cells but was exclusively unmethylated in a ynl024cΔ strain, and over-expression of Ynl024c caused a dramatic increase in Lys390 methylation, with trimethylation becoming the predominant state. Our results demonstrate that Ynl024c is the enzyme responsible for methylation of eEF1A at Lys390, and in accordance with prior naming of similar enzymes, we suggest that Ynl024c is renamed to Efm6 (Elongation factor MTase 6).
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Malecki, Jedrzej Mieczyslaw; Ho, Angela Yeuan Yen; Moen, Anders; Dahl, Helge-Andre & Falnes, Pål
(2015).
Human METTL20 is a mitochondrial lysine methyltransferase that targets the β subunit of electron transfer flavoprotein (ETFβ) and modulates its activity.
Journal of Biological Chemistry.
ISSN 0021-9258.
290(1),
p. 423–434.
doi:
10.1074/jbc.M114.614115.
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Haugsten, Ellen Margrethe; Malecki, Jedrzej M.; Bjørklund, Sunniva Maria Stordal; Olsnes, Sjur & Wesche, Jørgen
(2008).
Ubiquitination of fibroblast growth factor receptor 1 is required for its intracellular sorting but not for its endocytosis.
Molecular Biology of the Cell.
ISSN 1059-1524.
19,
p. 3390–3403.
doi:
10.1091/mbc.E07-12-1219.
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Wesche, Jørgen; Malecki, Jedrzej M.; Wiedlocha, Antoni; Skjerpen, Camilla Skiple; Claus, P & Olsnes, Sjur
(2006).
FGF-1 and FGF-2 require the cytosolic chaperone Hsp90 for translocation into the cytosol and the cell nucleus.
Journal of Biological Chemistry.
ISSN 0021-9258.
281,
p. 11405–11412.
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Wesche, Jørgen; Malecki, Jedrzej M.; Wiedlocha, Antoni; Ehsani, Maryam; Nilsen, Trine & Marcinkowska, Ewa
[Show all 7 contributors for this article]
(2005).
Two nuclear localization signals required for transport from the cytosol to the, nucleus of externally added FGF-1 translocated into cells.
Biochemistry (Moscow).
ISSN 0006-2979.
44.
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Malecki, Jedrzej M.; Wiedlocha, Antoni; Wesche, Jarlis & Olsnes, Sjur
(2002).
Vesicle transmembrane potential is required for translocation to cytosol externally added FGF-1.
EMBO Journal.
ISSN 0261-4189.
21,
p. 4480–4490.
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Published
Jan. 25, 2016 2:49 PM
- Last modified
Feb. 14, 2024 2:00 PM
