dc.contributor.author | Drazic, Adrian | |
dc.contributor.author | Timmerman, Evy | |
dc.contributor.author | Kajan, Ulrike | |
dc.contributor.author | Marie, Michaël Bruno Eric | |
dc.contributor.author | Varland, Sylvia | |
dc.contributor.author | Impens, Francis | |
dc.contributor.author | Gevaert, Kris | |
dc.contributor.author | Arnesen, Thomas | |
dc.date.accessioned | 2022-06-15T11:53:04Z | |
dc.date.available | 2022-06-15T11:53:04Z | |
dc.date.created | 2022-02-02T19:50:22Z | |
dc.date.issued | 2022 | |
dc.identifier.issn | 0022-2836 | |
dc.identifier.uri | https://hdl.handle.net/11250/2998877 | |
dc.description.abstract | Actin is a hallmark protein of the cytoskeleton in eukaryotic cells, affecting a range of cellular functions. Actin dynamics is regulated through a myriad of actin-binding proteins and post-translational modifications. The mammalian actin family consists of six different isoforms, which vary slightly in their N-terminal (Nt) sequences. During and after synthesis, actins undergo an intricate Nt-processing that yields mature actin isoforms. The ubiquitously expressed cytoplasmic β-actin is Nt-acetylated by N-alpha acetyltransferase 80 (NAA80) yielding the Nt-sequence Ac-DDDI-. In addition, β-actin was also reported to be Nt-arginylated by arginyltransferase 1 (ATE1) after further peptidase-mediated processing, yielding RDDI-. To characterize in detail the Nt-processing of actin, we used state-of-the-art proteomics. To estimate the relative cellular levels of Nt-modified proteoforms of actin, we employed NAA80-lacking cells, in which actin was not Nt-acetylated. We found that targeted proteomics is superior to a commercially available antibody previously used to analyze Nt-arginylation of β-actin. Significantly, despite the use of sensitive mass spectrometry-based techniques, we could not confirm the existence of the previously claimed Nt-arginylated β-actin (RDDI-) in either wildtype or NAA80-lacking cells. A very minor level of Nt-arginylation of the initially cleaved β-actin (DDDI-) could be identified, but only in NAA80-lacking cells, not in wildtype cells. We also identified small fractions of cleaved and unmodified β-actin (DDI-) as well as cleaved and Nt-acetylated β-actin (Ac-DDI-). In sum, we show that the multi-step Nt-maturation of β-actin is terminated by NAA80, which Nt-acetylates the exposed Nt-Asp residues, in the virtual absence of previously claimed Nt-arginylation. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | Elsevier | en_US |
dc.relation.uri | https://www.sciencedirect.com/science/article/pii/S0022283621006343?via%3Dihub#s0095 | |
dc.rights | Navngivelse 4.0 Internasjonal | * |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/deed.no | * |
dc.title | The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1 | en_US |
dc.type | Journal article | en_US |
dc.type | Peer reviewed | en_US |
dc.description.version | publishedVersion | en_US |
dc.rights.holder | Copyright 2021 The Author(s) | en_US |
dc.source.articlenumber | 167397 | en_US |
cristin.ispublished | true | |
cristin.fulltext | original | |
cristin.qualitycode | 1 | |
dc.identifier.doi | 10.1016/j.jmb.2021.167397 | |
dc.identifier.cristin | 1997158 | |
dc.source.journal | Journal of Molecular Biology (JMB) | en_US |
dc.relation.project | Norges forskningsråd: 249843 | en_US |
dc.relation.project | ERC-European Research Council: 772039 | en_US |
dc.relation.project | Helse Vest RHF: F-12540 | en_US |
dc.relation.project | Kreftforeningen: 171752—PR-2009-0222 | en_US |
dc.identifier.citation | Journal of Molecular Biology. 2022, 434 (2), 167397. | en_US |
dc.source.volume | 434 | en_US |
dc.source.issue | 2 | en_US |