vinayka

• 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. Full text in Research Archive Show summary

  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.

• Aileni, Vinay Kumar; Davydova, Erna; Moen, Anders & Falnes, Pål (2017). A System for Enzymatic Lysine Methylation in a Desired Sequence Context. ACS Omega. 2(2), p. 462–469. doi: 10.1021/acsomega.6b00486. Full text in Research Archive Show summary

  A number of lysine-specific methyltransferases (KMTs) are responsible for the post-translational modification of cellular proteins on lysine residues. Most KMTs typically recognize specific motifs in unstructured, short peptide sequences. However, we have recently discovered a novel KMT that appeared to have a more relaxed sequence specificity, namely, valosin-containing protein (VCP)-KMT, which trimethylates Lys-315 in the molecular chaperone VCP. On the basis of this, here, we explored the possibility of using the VCP-KMT/VCP system to obtain specific lysine methylation of desired sequences grafted onto a VCP-derived scaffold. We generated VCP-derived proteins in which three amino acid residues on each side of Lys-315 had been replaced by various sequences representing lysine methylation sites in histone H3. We found that all of these chimeric proteins were subject to efficient VCP-KMT-mediated methylation in vitro, and methylation was also observed in mammalian cells. Thus, we here describe a versatile system for introducing lysine methylation into a desired peptide sequence, and the approach should be readily expandable for generating combinatorial libraries of methylated sequences.

• Fusser, Markus; Kernstock, Stefan; Aileni, Vinay Kumar; Jacobsen, Wolfgang Mathias; Falnes, Pål & Klungland, Arne (2015). Lysine methylation of the valosin-containing protein (VCP) is dispensable for development and survival of mice. PLOS ONE. 10(11). doi: 10.1371/journal.pone.0141472. Full text in Research Archive

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• Aileni, Vinay Kumar; Davydova, Erna; Kernstock, Jens Stefan; Moen, Anders & Falnes, Pål (2016). VCP-KMT (METTL21D)-mediated lysine methylation of histone H3 peptide sequences grafted onto a VCP-derived scaffold. Full text in Research Archive

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