title: Identification and functional characterization of N-terminally acetylated proteins in Drosophila melanogaster.
abstract: Protein modifications play a major role for most biological processes in living organisms. Amino-terminal acetylation of proteins is a common modification found throughout the tree of life: the N-terminus of a nascent polypeptide chain becomes co-translationally acetylated, often after the removal of the initiating methionine residue. While the enzymes and protein complexes involved in these processes have been extensively studied, only little is known about the biological function of such N-terminal modification events. To identify common principles of N-terminal acetylation, we analyzed the amino-terminal peptides from proteins extracted from Drosophila Kc167 cells. We detected more than 1,200 mature protein N-termini and could show that N-terminal acetylation occurs in insects with a similar frequency as in humans. As the sole true determinant for N-terminal acetylation we could extract the (X)PX rule that indicates the prevention of acetylation under all circumstances. We could show that this rule can be used to genetically engineer a protein to study the biological relevance of the presence or absence of an acetyl group, thereby generating a generic assay to probe the functional importance of N-terminal acetylation. We applied the assay by expressing mutated proteins as transgenes in cell lines and in flies. Here, we present a straightforward strategy to systematically study the functional relevance of N-terminal acetylations in cells and whole organisms. Since the (X)PX rule seems to be of general validity in lower as well as higher eukaryotes, we propose that it can be used to study the function of N-terminal acetylation in all species.
authors.affiliation: Center for Model Organism Proteomes, University of Zurich, Switzerland. sandra.goetze@molbio.uzh.ch
authors.forename: Sandra
authors.initials: S
authors.lastname: Goetze
authors.forename: Ermir
authors.initials: E
authors.lastname: Qeli
authors.forename: Christian
authors.initials: C
authors.lastname: Mosimann
authors.forename: An
authors.initials: A
authors.lastname: Staes
authors.forename: Bertran
authors.initials: B
authors.lastname: Gerrits
authors.forename: Bernd
authors.initials: B
authors.lastname: Roschitzki
authors.forename: Sonali
authors.initials: S
authors.lastname: Mohanty
authors.forename: Eva M
authors.initials: EM
authors.lastname: Niederer
authors.forename: Endre
authors.initials: E
authors.lastname: Laczko
authors.forename: Evy
authors.initials: E
authors.lastname: Timmerman
authors.forename: Vinzenz
authors.initials: V
authors.lastname: Lange
authors.forename: Ernst
authors.initials: E
authors.lastname: Hafen
authors.forename: Ruedi
authors.initials: R
authors.lastname: Aebersold
authors.forename: Joël
authors.initials: J
authors.lastname: Vandekerckhove
authors.forename: Konrad
authors.initials: K
authors.lastname: Basler
authors.forename: Christian H
authors.initials: CH
authors.lastname: Ahrens
authors.forename: Kris
authors.initials: K
authors.lastname: Gevaert
authors.forename: Erich
authors.initials: E
authors.lastname: Brunner
mesh_terms: D000107:Acetylation; D000409:Alanine / Q000235:genetics / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites / Q000235:genetics; D015153:Blotting, Western; D002262:Carboxy-Lyases / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D030562:Databases, Protein; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D009154:Mutation; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D012694:Serine / Q000235:genetics / Q000378:metabolism; D013912:Threonine / Q000235:genetics / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
chemical_list: D029721:Drosophila Proteins; C508978:Hyx protein, Drosophila; D013912:Threonine; D012694:Serine; D002262:Carboxy-Lyases; D000409:Alanine