A novel mouse model for inhibition of DOHH-mediated hypusine modification reveals a crucial function in embryonic development, proliferation and oncogenic transformation.
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.
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany.
Henning
H
Sievert
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany. Heinrich Pette Institute, Leibniz Institute for Experimental Virology, 20251 Hamburg, Germany.
Nora
N
Pällmann
Center for Molecular Neurobiology (ZMNH), University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, Germany.
Katharine K
KK
Miller
Center for Molecular Neurobiology (ZMNH), University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, Germany.
Irm
I
Hermans-Borgmeyer
Department of Medical Biochemistry and Molecular Biology, University of Greifswald, 17475 Greifswald, Germany.
Simone
S
Venz
Institute of Molecular Life Sciences, University of Zurich, 8057 Zurich, Switzerland. Division of Hematology, University Hospital Zurich, 8091 Zurich, Switzerland.
Ataman
A
Sendoel
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany.
Michael
M
Preukschas
Center for Molecular Neurobiology (ZMNH), University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, Germany.
Michaela
M
Schweizer
Division of Hematology, University Hospital Zurich, 8091 Zurich, Switzerland.
Steffen
S
Boettcher
Center for Molecular Neurobiology (ZMNH), University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, Germany.
P Christoph
PC
Janiesch
Department of Clinical Chemistry, University Hospital of Cologne, 50924 Cologne, Germany.
Thomas
T
Streichert
Department of Medical Biochemistry and Molecular Biology, University of Greifswald, 17475 Greifswald, Germany.
Reinhard
R
Walther
Institute of Molecular Life Sciences, University of Zurich, 8057 Zurich, Switzerland.
Michael O
MO
Hengartner
Division of Hematology, University Hospital Zurich, 8091 Zurich, Switzerland.
Markus G
MG
Manz
Clinic for Internal Medicine IV, Hematology and Oncology, University Hospital of the RWTH Aachen, 52074 Aachen, Germany.
Tim H
TH
Brümmendorf
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany.
Carsten
C
Bokemeyer
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany.
Melanie
M
Braig
Heinrich Pette Institute, Leibniz Institute for Experimental Virology, 20251 Hamburg, Germany.
Joachim
J
Hauber
Center for Molecular Neurobiology (ZMNH), University Medical Center Hamburg-Eppendorf (UKE), 20251 Hamburg, Germany.
Kent E
KE
Duncan
Department of Oncology, Hematology and Bone Marrow Transplantation with Section Pneumology, Hubertus Wald-Tumor Zentrum, University Hospital Eppendorf, 20246 Hamburg, Germany. Division of Hematology, University Hospital Zurich, 8091 Zurich, Switzerland. stefan.balabanov@usz.ch.
Stefan
S
Balabanov
D016475:3T3 Cells; D000483:Alleles; D000818:Animals; D017173:Caenorhabditis elegans; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic / Q000378:metabolism* / Q000473:pathology*; D016922:Cellular Senescence; D004195:Disease Models, Animal; D020964:Embryo Loss / Q000378:metabolism / Q000473:pathology; D047108:Embryonic Development; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D055786:Gene Knockout Techniques; D006900:Hydroxylation; D008239:Lysine / Q000031:analogs & derivatives* / Q000378:metabolism; D051379:Mice; D006899:Mixed Function Oxygenases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism; D010641:Phenotype; D014176:Protein Biosynthesis; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D018631:ras Proteins / Q000378:metabolism; D000097574:Eukaryotic Translation Initiation Factor 5A
D008239:Lysine; D006899:Mixed Function Oxygenases; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010448:Peptide Initiation Factors; D016271:Proto-Oncogene Proteins c-myc; D016601:RNA-Binding Proteins; D018631:ras Proteins; D000097574:Eukaryotic Translation Initiation Factor 5A; C100028:hypusine; C047991:deoxyhypusine hydroxylase; C063507:deoxyhypusine synthase
Cancer; Hypusine modification; Mouse models; Translational control