Protein-protein-interaction network organization of the hypusine modification system.
Hypusine modification of eukaryotic initiation factor 5A (eIF-5A) represents a unique and highly specific post-translational modification with regulatory functions in cancer, diabetes, and infectious diseases. However, the specific cellular pathways that are influenced by the hypusine modification remain largely unknown. To globally characterize eIF-5A and hypusine-dependent pathways, we used an approach that combines large-scale bioreactor cell culture with tandem affinity purification and mass spectrometry: "bioreactor-TAP-MS/MS." By applying this approach systematically to all four components of the hypusine modification system (eIF-5A1, eIF-5A2, DHS, and DOHH), we identified 248 interacting proteins as components of the cellular hypusine network, with diverse functions including regulation of translation, mRNA processing, DNA replication, and cell cycle regulation. Network analysis of this data set enabled us to provide a comprehensive overview of the protein-protein interaction landscape of the hypusine modification system. In addition, we validated the interaction of eIF-5A with some of the newly identified associated proteins in more detail. Our analysis has revealed numerous novel interactions, and thus provides a valuable resource for understanding how this crucial homeostatic signaling pathway affects different cellular functions.
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, and Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology, Hamburg, Germany.
Henning
H
Sievert
Simone
S
Venz
Oscar
O
Platas-Barradas
Vishnu M
VM
Dhople
Martin
M
Schaletzky
Claus-Henning
CH
Nagel
Melanie
M
Braig
Michael
M
Preukschas
Nora
N
Pällmann
Carsten
C
Bokemeyer
Tim H
TH
Brümmendorf
Ralf
R
Pörtner
Reinhard
R
Walther
Kent E
KE
Duncan
Joachim
J
Hauber
Stefan
S
Balabanov
D000818:Animals; D019295:Computational Biology; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D006899:Mixed Function Oxygenases / Q000378:metabolism; D057146:Multivesicular Bodies / Q000378:metabolism; D041681:NIH 3T3 Cells; D009687:Nuclear Proteins / Q000378:metabolism; D000090243:Nucleophosmin; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism; D060066:Protein Interaction Maps; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results; D012269:Ribosomal Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D000097574:Eukaryotic Translation Initiation Factor 5A
D004268:DNA-Binding Proteins; D008239:Lysine; D006899:Mixed Function Oxygenases; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010446:Peptide Fragments; D010448:Peptide Initiation Factors; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A; C100028:hypusine; C047991:deoxyhypusine hydroxylase; C063507:deoxyhypusine synthase