title: Mutations in CDC45, Encoding an Essential Component of the Pre-initiation Complex, Cause Meier-Gorlin Syndrome and Craniosynostosis.
abstract: DNA replication precisely duplicates the genome to ensure stable inheritance of genetic information. Impaired licensing of origins of replication during the G1 phase of the cell cycle has been implicated in Meier-Gorlin syndrome (MGS), a disorder defined by the triad of short stature, microtia, and a/hypoplastic patellae. Biallelic partial loss-of-function mutations in multiple components of the pre-replication complex (preRC; ORC1, ORC4, ORC6, CDT1, or CDC6) as well as de novo stabilizing mutations in the licensing inhibitor, GMNN, cause MGS. Here we report the identification of mutations in CDC45 in 15 affected individuals from 12 families with MGS and/or craniosynostosis. CDC45 encodes a component of both the pre-initiation (preIC) and CMG helicase complexes, required for initiation of DNA replication origin firing and ongoing DNA synthesis during S-phase itself, respectively, and hence is functionally distinct from previously identified MGS-associated genes. The phenotypes of affected individuals range from syndromic coronal craniosynostosis to severe growth restriction, fulfilling diagnostic criteria for Meier-Gorlin syndrome. All mutations identified were biallelic and included synonymous mutations altering splicing of physiological CDC45 transcripts, as well as amino acid substitutions expected to result in partial loss of function. Functionally, mutations reduce levels of full-length transcripts and protein in subject cells, consistent with partial loss of CDC45 function and a predicted limited rate of DNA replication and cell proliferation. Our findings therefore implicate the preIC as an additional protein complex involved in the etiology of MGS and connect the core cellular machinery of genome replication with growth, chondrogenesis, and cranial suture homeostasis.
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Aimee L
authors.initials: AL
authors.lastname: Fenwick
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK; Department of Oncology, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK.
authors.forename: Maciej
authors.initials: M
authors.lastname: Kliszczak
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Fay
authors.initials: F
authors.lastname: Cooper
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Jennie
authors.initials: J
authors.lastname: Murray
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Luis
authors.initials: L
authors.lastname: Sanchez-Pulido
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Stephen R F
authors.initials: SR
authors.lastname: Twigg
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Goriely
authors.affiliation: Computational Biology Research Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: McGowan
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Kerry A
authors.initials: KA
authors.lastname: Miller
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK.
authors.forename: Indira B
authors.initials: IB
authors.lastname: Taylor
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Clare
authors.initials: C
authors.lastname: Logan
authors.affiliation: Department of Medical Genetics, Mersin University, Mersin, 33343 Cukurova, Turkey.
authors.forename: Sevcan
authors.initials: S
authors.lastname: Bozdogan
authors.affiliation: Department of Clinical Genetics, Christian Medical College and Hospital, Vellore, Tamil Nadu 632004, India.
authors.forename: Sumita
authors.initials: S
authors.lastname: Danda
authors.affiliation: Genetic Health Service NZ-South Island Hub, Christchurch Hospital, Christchurch, Canterbury 8140, New Zealand.
authors.forename: Joanne
authors.initials: J
authors.lastname: Dixon
authors.affiliation: Children's Hospital, Ain Shams University, Cairo 11566, Egypt.
authors.forename: Solaf M
authors.initials: SM
authors.lastname: Elsayed
authors.affiliation: Children's Hospital, Ain Shams University, Cairo 11566, Egypt.
authors.forename: Ezzat
authors.initials: E
authors.lastname: Elsobky
authors.affiliation: North East Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, Great Ormond Street Hospital, London WC1N 3JH, UK.
authors.forename: Alice
authors.initials: A
authors.lastname: Gardham
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
authors.forename: Mariette J V
authors.initials: MJ
authors.lastname: Hoffer
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
authors.forename: Marije
authors.initials: M
authors.lastname: Koopmans
authors.affiliation: Clinical Genetics, The Children's Hospital of Philadelphia, 34th & Civic Center Boulevard, Philadelphia, PA 19104, USA.
authors.forename: Donna M
authors.initials: DM
authors.lastname: McDonald-McGinn
authors.affiliation: Department of Clinical Genetics, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
authors.forename: Gijs W E
authors.initials: GW
authors.lastname: Santen
authors.affiliation: Victorian Clinical Genetics Services, Murdoch Children's Research Institute, University of Melbourne, Melbourne, VIC 3052, Australia.
authors.forename: Ravi
authors.initials: R
authors.lastname: Savarirayan
authors.affiliation: Department of Physiology, Faculty of Medicine, University of Kelaniya, Ragama, Gampaha GQ 11010, Sri Lanka.
authors.forename: Deepthi
authors.initials: D
authors.lastname: de Silva
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, 9000 Ghent, Belgium.
authors.forename: Olivier
authors.initials: O
authors.lastname: Vanakker
authors.affiliation: Craniofacial Unit, Department of Plastic and Reconstructive Surgery, Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, Oxford OX3 9DU, UK.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Wall
authors.affiliation: North East Thames Regional Genetics Service, Great Ormond Street Hospital for Children NHS Foundation Trust, Great Ormond Street Hospital, London WC1N 3JH, UK.
authors.forename: Louise C
authors.initials: LC
authors.lastname: Wilson
authors.affiliation: Genetic Diagnosis Center, Adana Numune Training and Research Hospital, Cukurova, Adana, 01170, Turkey.
authors.forename: Ozge Ozalp
authors.initials: OO
authors.lastname: Yuregir
authors.affiliation: Clinical Genetics, The Children's Hospital of Philadelphia, 34th & Civic Center Boulevard, Philadelphia, PA 19104, USA.
authors.forename: Elaine H
authors.initials: EH
authors.lastname: Zackai
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Chris P
authors.initials: CP
authors.lastname: Ponting
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Jackson
authors.affiliation: Clinical Genetics Group, MRC Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, UK; Craniofacial Unit, Department of Plastic and Reconstructive Surgery, Oxford University Hospitals NHS Foundation Trust, John Radcliffe Hospital, Oxford OX3 9DU, UK.
authors.forename: Andrew O M
authors.initials: AO
authors.lastname: Wilkie
authors.affiliation: Department of Oncology, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DS, UK. Electronic address: wojciech.niedzwiedz@imm.ox.ac.uk.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Niedzwiedz
authors.affiliation: MRC Human Genetics Unit, IGMM, University of Edinburgh, Edinburgh EH4 2XU, UK; Department of Pathology, Dunedin School of Medicine, University of Otago, Dunedin, Otago 9016, New Zealand. Electronic address: louise.bicknell@otago.ac.nz.
authors.forename: Louise S
authors.initials: LS
authors.lastname: Bicknell
mesh_terms: D000293:Adolescent; D000328:Adult; D000483:Alleles; D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D000650:Amnion / Q000166:cytology; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D065817:Congenital Microtia / Q000235:genetics*; D003398:Craniosynostoses / Q000235:genetics*; D004252:DNA Mutational Analysis; D004261:DNA Replication; D059472:Exome / Q000235:genetics; D005091:Exons / Q000235:genetics; D005260:Female; D056726:Genetic Association Studies; D006130:Growth Disorders / Q000235:genetics*; D006801:Humans; D008297:Male; D008844:Micrognathism / Q000235:genetics*; D008958:Models, Molecular; D009154:Mutation; D010329:Patella / Q000002:abnormalities*; D011487:Protein Conformation; D013577:Syndrome; D055815:Young Adult
chemical_list: C113571:CDC45 protein, human; D018797:Cell Cycle Proteins