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.

Subjects

Animals

Computational Biology

DNA-Binding Proteins

Humans

Lysine

Mass Spectrometry

Mice

Mixed Function Oxygenases

Multivesicular Bodies

NIH 3T3 Cells

Nuclear Proteins

Oxidoreductases Acting on CH-NH Group Donors

Peptide Fragments

Peptide Initiation Factors

Protein Transport

RNA-Binding Proteins

Recombinant Fusion Proteins

Reproducibility of Results

Ribosomal Proteins

Subcellular Fractions

Protein Interaction Maps

Protein Processing, Post-Translational

DDC Class

570: Biowissenschaften, Biologie

610: Medizin

Lizenz

https://creativecommons.org/licenses/by/4.0/

Name

1289.full.pdf

Size

4.51 MB

Format

Adobe PDF

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Protein-protein-interaction network organization of the hypusine modification system