Víctor Faundes1,2, Martin D Jennings3,4, Siobhan Crilly5, Sarah Legraie1, Sarah E Withers5, Sara Cuvertino1, Sally J Davies6, Andrew G L Douglas7,8, Andrew E Fry6,9, Victoria Harrison7, Jeanne Amiel10,11,12, Daphné Lehalle10, William G Newman1,13, Patricia Newkirk14, Judith Ranells14, Miranda Splitt15, Laura A Cross16,17, Carol J Saunders18,19,20, Bonnie R Sullivan16,17, Jorge L Granadillo21, Christopher T Gordon11,12, Paul R Kasher22,23, Graham D Pavitt24,25, Siddharth Banka26,27. 1. Division of Evolution & Genomic Sciences, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. 2. Laboratorio de Genética y Enfermedades Metabólicas, Instituto de Nutrición y Tecnología de los Alimentos (INTA), Universidad de Chile, Santiago, Chile. 3. Division of Molecular and Cellular Function, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. 4. Manchester Academic Health Science Centre, University of Manchester, Manchester, UK. 5. Division of Neuroscience & Experimental Psychology, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. 6. Institute of Medical Genetics, University Hospital of Wales, Cardiff, UK. 7. Wessex Clinical Genetics Service, Princess Anne Hospital, Southampton, UK. 8. Human Development and Health, Faculty of Medicine, University of Southampton, Southampton General Hospital, Southampton, UK. 9. Division of Cancer and Genetics, School of Medicine, Cardiff University, Cardiff, UK. 10. Department of Genetics, AP-HP, Hôpital Necker Enfants Malades, Paris, France. 11. 1Laboratory of Embryology and Genetics of Human Malformations, INSERM UMR 1163, Institut Imagine, Paris, France. 12. Paris Descartes-Sorbonne Paris Cité University, Institut Imagine, Paris, France. 13. Manchester Centre for Genomic Medicine, St Mary's Hospital, Manchester University NHS Foundation Trust, Health Innovation Manchester, Manchester, UK. 14. Division of Genetics and Metabolism, Department of Pediatrics, University of South Florida, Tampa, FL, UK. 15. Northern Genetics Service, Institute of Genetic Medicine, Newcastle upon Tyne, UK. 16. Division of Clinical Genetics, Children's Mercy, Kansas City, MO, USA. 17. Department of Pediatrics, University of Missour-Kansas City, Kansas City, MO, USA. 18. Center for Pediatric Genomic Medicine Children's Mercy, Kansas City, MO, USA. 19. School of Medicine, University of Missouri-Kansas City, Kansas City, MO, USA. 20. Department of Pathology and Laboratory Medicine, Children's Mercy, Kansas City, MO, USA. 21. Division of Genetics and Genomic Medicine, Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, USA. 22. Manchester Academic Health Science Centre, University of Manchester, Manchester, UK. paul.kasher@manchester.ac.uk. 23. Division of Neuroscience & Experimental Psychology, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. paul.kasher@manchester.ac.uk. 24. Division of Molecular and Cellular Function, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. graham.pavitt@manchester.ac.uk. 25. Manchester Academic Health Science Centre, University of Manchester, Manchester, UK. graham.pavitt@manchester.ac.uk. 26. Division of Evolution & Genomic Sciences, School of Biological Sciences, Faculty of Biology, Medicine and Health, University of Manchester, Manchester, UK. siddharth.banka@manchester.ac.uk. 27. Manchester Centre for Genomic Medicine, St Mary's Hospital, Manchester University NHS Foundation Trust, Health Innovation Manchester, Manchester, UK. siddharth.banka@manchester.ac.uk.
Abstract
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.
The structure of proline prevents it from adopting an optimal position for rapid protein synthesis. pan class="Chemical">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.
Authors: Susan Zeesman; Susanne Kjaergaard; Hanne D Hove; Maria Kirchhoff; Jadd M Stevens; Małgorzata J M Nowaczyk Journal: Am J Med Genet A Date: 2012-06-29 Impact factor: 2.802
Authors: Anthony A Philippakis; Danielle R Azzariti; Sergi Beltran; Anthony J Brookes; Catherine A Brownstein; Michael Brudno; Han G Brunner; Orion J Buske; Knox Carey; Cassie Doll; Sergiu Dumitriu; Stephanie O M Dyke; Johan T den Dunnen; Helen V Firth; Richard A Gibbs; Marta Girdea; Michael Gonzalez; Melissa A Haendel; Ada Hamosh; Ingrid A Holm; Lijia Huang; Matthew E Hurles; Ben Hutton; Joel B Krier; Andriy Misyura; Christopher J Mungall; Justin Paschall; Benedict Paten; Peter N Robinson; François Schiettecatte; Nara L Sobreira; Ganesh J Swaminathan; Peter E Taschner; Sharon F Terry; Nicole L Washington; Stephan Züchner; Kym M Boycott; Heidi L Rehm Journal: Hum Mutat Date: 2015-10 Impact factor: 4.878
Authors: D Lehalle; D Wieczorek; R M Zechi-Ceide; M R Passos-Bueno; S Lyonnet; J Amiel; C T Gordon Journal: Clin Genet Date: 2015-05-01 Impact factor: 4.438
Authors: Erik Gutierrez; Byung-Sik Shin; Christopher J Woolstenhulme; Joo-Ran Kim; Preeti Saini; Allen R Buskirk; Thomas E Dever Journal: Mol Cell Date: 2013-05-30 Impact factor: 17.970
Authors: Sara Cuvertino; Helen M Stuart; Kate E Chandler; Neil A Roberts; Ruth Armstrong; Laura Bernardini; Sanjeev Bhaskar; Bert Callewaert; Jill Clayton-Smith; Cristina Hernando Davalillo; Charu Deshpande; Koenraad Devriendt; Maria C Digilio; Abhijit Dixit; Matthew Edwards; Jan M Friedman; Antonio Gonzalez-Meneses; Shelagh Joss; Bronwyn Kerr; Anne Katrin Lampe; Sylvie Langlois; Rachel Lennon; Philippe Loget; David Y T Ma; Ruth McGowan; Maryse Des Medt; James O'Sullivan; Sylvie Odent; Michael J Parker; Céline Pebrel-Richard; Florence Petit; Zornitza Stark; Sylvia Stockler-Ipsiroglu; Sigrid Tinschert; Pradeep Vasudevan; Olaya Villa; Susan M White; Farah R Zahir; Adrian S Woolf; Siddharth Banka Journal: Am J Hum Genet Date: 2017-12-07 Impact factor: 11.025
Authors: Emanuela Santini; Thu N Huynh; Andrew F MacAskill; Adam G Carter; Philippe Pierre; Davide Ruggero; Hanoch Kaphzan; Eric Klann Journal: Nature Date: 2012-12-23 Impact factor: 49.962
Authors: Emily Connell; Gwenaelle Le Gall; Matthew G Pontifex; Saber Sami; John F Cryan; Gerard Clarke; Michael Müller; David Vauzour Journal: Mol Neurodegener Date: 2022-06-17 Impact factor: 18.879
Authors: Ebelechukwu C Nwokoye; Eiman AlNaseem; Robert A Crawford; Lydia M Castelli; Martin D Jennings; Christopher J Kershaw; Graham D Pavitt Journal: Sci Rep Date: 2021-06-29 Impact factor: 4.379