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Updated: Apr 29, 2026
Inducible and Reversible Dominant-negative DN Protein Inhibition
Published on: January 7, 2019
A novel mouse model for inhibition of DOHH-mediated hypusine modification reveals a crucial function in embryonic
Henning Sievert1, Nora Pällmann2, Katharine K Miller3
1Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany.
Abstract:
The central importance of translational control by post-translational modification has spurred major interest in regulatory pathways that control translation. One such pathway uniquely adds hypusine to eukaryotic initiation factor 5A (eIF5A), and thereby affects protein synthesis and, subsequently, cellular proliferation through an unknown mechanism. Using a novel conditional knockout mouse model and a Caenorhabditis elegans knockout model, we found an evolutionarily conserved role for the DOHH-mediated second step of hypusine synthesis in early embryonic development. At the cellular level, we observed reduced proliferation and induction of senescence in 3T3 Dohh-/- cells as well as reduced capability for malignant transformation. Furthermore, mass spectrometry showed that deletion of DOHH results in an unexpected complete loss of hypusine modification. Our results provide new biological insight into the physiological roles of the second step of the hypusination of eIF5A. Moreover, the conditional mouse model presented here provides a powerful tool for manipulating hypusine modification in a temporal and spatial manner, to analyse both how this unique modification normally functions in vivo as well as how it contributes to different pathological conditions.
Insights
The DOHH enzyme
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Post-translational modification regulates translation, impacting protein synthesis and cellular proliferation.
- Hypusination of eukaryotic initiation factor 5A (eIF5A) is a unique regulatory pathway.
- The precise physiological roles of hypusination, particularly the second step, remain largely unknown.
Purpose of the Study:
- To investigate the evolutionary conserved role of the DOHH-mediated second step of hypusine synthesis.
- To elucidate the cellular and physiological consequences of DOHH deletion.
- To establish a conditional mouse model for studying hypusination in vivo.
Main Methods:
- Generation and utilization of conditional knockout mouse and Caenorhabditis elegans models.
- Cellular proliferation and senescence assays in 3T3 Dohh-/- cells.
- Mass spectrometry to analyze hypusine modification levels.
Main Results:
- DOHH plays a conserved role in early embryonic development.
- DOHH deletion leads to reduced cellular proliferation, induced senescence, and impaired malignant transformation.
- Complete loss of hypusine modification was observed upon DOHH deletion.
- The conditional mouse model allows for temporal and spatial manipulation of hypusination.
Conclusions:
- The second step of hypusination, mediated by DOHH, is crucial for normal embryonic development and cellular homeostasis.
- DOHH is essential for eIF5A hypusination, impacting protein synthesis and cell proliferation.
- The developed mouse model is a valuable tool for future research into hypusination's role in health and disease.
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