TY - JOUR
T1 - Impaired eIF5A function causes a Mendelian disorder that is partially rescued in model systems by spermidine
AU - Faundes, Víctor
AU - Jennings, Martin D.
AU - Crilly, Siobhan
AU - Legraie, Sarah
AU - Withers, Sarah E.
AU - Cuvertino, Sara
AU - Davies, Sally J.
AU - Douglas, Andrew G.L.
AU - Fry, Andrew E.
AU - Harrison, Victoria
AU - Amiel, Jeanne
AU - Lehalle, Daphné
AU - Newman, William G.
AU - Newkirk, Patricia
AU - Ranells, Judith
AU - Splitt, Miranda
AU - Cross, Laura A.
AU - Saunders, Carol J.
AU - Sullivan, Bonnie R.
AU - Granadillo, Jorge L.
AU - Gordon, Christopher T.
AU - Kasher, Paul R.
AU - Pavitt, Graham D.
AU - Banka, Siddharth
N1 - Funding Information:
We are thankful to all the individuals and their families for taking part in the study. We thank Tom Dever (National Institutes of Health, USA) and Beth Grayhack (University of Rochester Medical Center, USA) for kind gifts of plasmids and yeast strains used in this study, as well as Martin Pool (University of Manchester, UK) for the gift of antibodies to ribosomal proteins. We are thankful to the Deciphering Developmental Disorders (DDD) study for the invaluable collaboration. The DDD Study (Cambridge South REC approval 10/H0305/83 and the Republic of Ireland REC GEN/284/12) presents independent research commissioned by the Health Innovation Challenge Fund (grant number HICF-1009-003), a parallel funding partnership between the Wellcome Trust and the Department of Health and the Wellcome Trust Sanger Institute (grant number WT098051). The views expressed in this publication are those of the author(s) and not necessarily those of the Wellcome Trust, BBSRC or the Department of Health. The research team acknowledges the support of the National Institute for Health Research, through the Comprehensive Clinical Research Network, UK. V.F. acknowledges to CONICYT, Chile’s National Commission for Scientific and Technological Research, for its scholarship support (grant number 72160007). V.F., W.G.N. and S.B. acknowledge to the Kabuki Research Fund at Manchester University NHS Foundation Trust. W.G.N. acknowledges support from Action Medical Research (GN2494), and the Manchester NIHR Biomedical Research Centre (IS-BRC-1215-20007). G.D.P. and M.D.J. acknowledge to Biotechnology and Biological Sciences Research Council (BBSRC), UK, for its financial support (grant BB/N014049/1). P.R.K and S. Crilly were supported by the Stroke Association (TSA LECT 2017/02) and the NC3Rs (NC/N002598/1). J.A. and C.T.G. were supported by the Agence Nationale de la Recherche (CranioRespiro project and ‘Investissements d’avenir' program (ANR-10-IAHU-01)) and MSDAvenir (Devo-Decode project).
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - The structure of proline prevents it from adopting an optimal position for rapid protein synthesis. Poly-proline-tract (PPT) associated ribosomal stalling is resolved by highly conserved eIF5A, the only protein to contain the amino acid hypusine. We show that de novo heterozygous EIF5A variants cause a disorder characterized by variable combinations of developmental delay, microcephaly, micrognathia and dysmorphism. Yeast growth assays, polysome profiling, total/hypusinated eIF5A levels and PPT-reporters studies reveal that the variants impair eIF5A function, reduce eIF5A-ribosome interactions and impair the synthesis of PPT-containing proteins. Supplementation with 1 mM spermidine partially corrects the yeast growth defects, improves the polysome profiles and restores expression of PPT reporters. In zebrafish, knockdown eif5a partly recapitulates the human phenotype that can be rescued with 1 µM spermidine supplementation. In summary, we uncover the role of eIF5A in human development and disease, demonstrate the mechanistic complexity of EIF5A-related disorder and raise possibilities for its treatment.
AB - The structure of proline prevents it from adopting an optimal position for rapid protein synthesis. Poly-proline-tract (PPT) associated ribosomal stalling is resolved by highly conserved eIF5A, the only protein to contain the amino acid hypusine. We show that de novo heterozygous EIF5A variants cause a disorder characterized by variable combinations of developmental delay, microcephaly, micrognathia and dysmorphism. Yeast growth assays, polysome profiling, total/hypusinated eIF5A levels and PPT-reporters studies reveal that the variants impair eIF5A function, reduce eIF5A-ribosome interactions and impair the synthesis of PPT-containing proteins. Supplementation with 1 mM spermidine partially corrects the yeast growth defects, improves the polysome profiles and restores expression of PPT reporters. In zebrafish, knockdown eif5a partly recapitulates the human phenotype that can be rescued with 1 µM spermidine supplementation. In summary, we uncover the role of eIF5A in human development and disease, demonstrate the mechanistic complexity of EIF5A-related disorder and raise possibilities for its treatment.
UR - http://www.scopus.com/inward/record.url?scp=85100582617&partnerID=8YFLogxK
U2 - 10.1038/s41467-021-21053-2
DO - 10.1038/s41467-021-21053-2
M3 - Article
C2 - 33547280
AN - SCOPUS:85100582617
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 833
ER -