title: Lysine Methylation of the Valosin-Containing Protein (VCP) Is Dispensable for Development and Survival of Mice.
abstract: Valosin-containing protein (VCP) is a homohexameric ATPase involved in a multitude cellular processes and it was recently shown that VCP is trimethylated at lysine 315 by the VCP lysine methyltransferase (VCPKMT). Here, we generated and validated a constitutive knockout mouse by targeting exon 1-4 of the Vcpkmt gene. We show that Vcpkmt is ubiquitously expressed in all tissues examined and confirm the sub-cellular localization to the cytoplasm. We show by (I) mass spectrometric analysis, (II) VCPKMT-mediated in vitro methylation of VCP in cell extracts and (III) immunostaining with a methylation specific antibody, that in Vcpkmt-/- mice the methylation of lysine 315 in VCP is completely abolished. In contrast, VCP is almost exclusively trimethylated in wild-type mice. Furthermore, we investigated the specificity of VCPKMT with in vitro methylation assays using as source of substrate protein extracts from Vcpkmt-/- mouse organs or three human Vcpkmt-/- cell lines. The results show that VCPKMT is a highly specific enzyme, and suggest that VCP is its sole substrate. The Vcpkmt-/- mice were viable, fertile and had no obvious pathological phenotype. Their body weight, life span and acute endurance capacity were comparable to wild-type controls. Overall the results show that VCPKMT is an enzyme required for methylation of K315 of VCP in vivo, but VCPKMT is not essential for development or survival under unstressed conditions.
authors.affiliation: Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Oslo, Norway.
authors.forename: Markus
authors.initials: M
authors.lastname: Fusser
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Norway.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kernstock
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Norway.
authors.forename: Vinay Kumar
authors.initials: VK
authors.lastname: Aileni
authors.affiliation: Glyconor Mass Spectrometry, Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Norway.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Egge-Jacobsen
authors.affiliation: Department of Biosciences, Faculty of Mathematics and Natural Sciences, University of Oslo, Oslo, Norway.
authors.forename: Pål Ø
authors.initials: PØ
authors.lastname: Falnes
authors.affiliation: Institute of Medical Microbiology, Oslo University Hospital, Rikshospitalet, Oslo, Norway.|Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.
authors.forename: Arne
authors.initials: A
authors.lastname: Klungland
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism*; D005260:Female; D005298:Fertility; D018507:Gene Expression Regulation, Developmental; D055786:Gene Knockout Techniques; D048788:Growth and Development; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008297:Male; D008745:Methylation; D008780:Methyltransferases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D010641:Phenotype; D013379:Substrate Specificity; D016019:Survival Analysis; D000074405:Valosin Containing Protein
chemical_list: D018797:Cell Cycle Proteins; D008780:Methyltransferases; C000608434:VCP lysine methyltransferase, mouse; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein; C000614179:Vcp protein, mouse; D008239:Lysine