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dc.contributor.authorDrazic, Adrian
dc.contributor.authorTimmerman, Evy
dc.contributor.authorKajan, Ulrike
dc.contributor.authorMarie, Michaël Bruno Eric
dc.contributor.authorVarland, Sylvia
dc.contributor.authorImpens, Francis
dc.contributor.authorGevaert, Kris
dc.contributor.authorArnesen, Thomas
dc.description.abstractActin 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.rightsNavngivelse 4.0 Internasjonal*
dc.titleThe Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1en_US
dc.typeJournal articleen_US
dc.typePeer revieweden_US
dc.rights.holderCopyright 2021 The Author(s)en_US
dc.source.journalJournal of Molecular Biology (JMB)en_US
dc.relation.projectNorges forskningsråd: 249843en_US
dc.relation.projectERC-European Research Council: 772039en_US
dc.relation.projectHelse Vest RHF: F-12540en_US
dc.relation.projectKreftforeningen: 171752—PR-2009-0222en_US
dc.identifier.citationJournal of Molecular Biology. 2022, 434 (2), 167397.en_US

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