NatF Contributes to an Evolutionary Shift in Protein N-Terminal Acetylation and Is Important for Normal Chromosome Segregation 英文参考文献.docVIP

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NatF Contributes to an Evolutionary Shift in Protein N-Terminal Acetylation and Is Important for Normal Chromosome Segregation 英文参考文献.doc

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NatF Contributes to an Evolutionary Shift in Protein N-Terminal Acetylation and Is Important for Normal Chromosome Segregation 英文参考文献

NatFContributestoanEvolutionaryShiftinProtein N-TerminalAcetylationandIsImportantforNormal ChromosomeSegregation PetraVanDamme1,2.,KristineHole3,4.,AnaPimenta-Marques5.,KennyHelsens1,2,Joe¨l Vandekerckhove1,2,RuiG.Martinho5,KrisGevaert1,2,ThomasArnesen3,6 * 1DepartmentofMedicalProteinResearch,GhentUniversity,Ghent,Belgium,2DepartmentofBiochemistry,GhentUniversity,Ghent,Belgium,3DepartmentofMolecular Biology, University of Bergen, Bergen, Norway, 4Department of Surgical Sciences, University of Bergen, Bergen, Norway, 5Instituto Gulbenkian de Cie?ncia, Oeiras, Portugal,6DepartmentofSurgery,HaukelandUniversityHospital,Bergen,Norway Abstract N-terminal acetylation (N-Ac) is a highly abundant eukaryotic protein modification. Proteomics revealed a significant increase in the occurrence of N-Ac from lower to higher eukaryotes, but evidence explaining the underlying molecular mechanism(s) is currently lacking. We first analysed protein N-termini and their acetylation degrees, suggesting that evolution of substrates is not a major cause for the evolutionary shift in N-Ac. Further, we investigated the presence of putative N-terminal acetyltransferases (NATs) in higher eukaryotes. The purified recombinant human and Drosophila homologuesofanovelNATcandidatewassubjectedtoinvitropeptidelibraryacetylationassays.Thisprovidedevidence foritsNATactivitytargetingMet-Lys-andotherMet-startingproteinN-termini,andtheenzymewastermedNaa60pandits activityNatF.ItsinvivoactivitywasinvestigatedbyectopicallyexpressinghumanNaa60pinyeastfollowedbyN-terminal COFRADIC analyses. hNaa60p acetylated distinct Met-starting yeast protein N-termini and increased general acetylation levels,therebyalteringyeastinvivoacetylationpatternstowardsthoseofhighereukaryotes.Further,itsactivityinhuman cellswasverifiedbyoverexpressionandknockdownofhNAA60followedbyN-terminalCOFRADIC.NatF’scellularimpact wasdemonstratedinDrosophilacellswhereNAA60knockdowninducedchromosomalsegregationdefects.Insummary, ourstudyrevealeda