Abstract
Objective The characterization of a novel large deletion in the galactose-1-phosphate uridyltransferase (GALT) gene accounting for the majority of disease alleles in Cypriot patients with classic galactosemia.
Methods DNA sequencing was used to identify the mutations followed by multiplex ligation-dependent probe amplification (MLPA) analysis in the cases suspected of harboring a deletion. In order to map the breakpoints of the novel deletion, a PCR walking approach was employed. A simple PCR assay was validated for diagnostic testing for the new deletion. Haplotype analysis was performed using microsatellite markers in the chromosomal region 9p. RT-PCR was used to study RNA expression in lymphoblastoid cell lines.
Results The new deletion spans a region of 8489 bp and eliminates all GALT exons as well as the non-translated sequences of the adjacent interleukin 11 receptor alpha (IL11RA) gene. In addition, the deletion is flanked by a 6 bp block of homologous sequence on either side suggesting that a single deletion event has occurred, probably mediated by a recombination mechanism. Microsatellite marker analysis revealed the existence of a common haplotype. The RNA expression studies showed a lack of IL11RA transcripts in patients homozygous for the deletion.
Conclusions We have identified and characterized a novel contiguous deletion which affects both the GALT enzyme and the IL11RA protein resulting in classic galactosemia with additional phenotypic abnormalities such as craniosynostosis, a feature that has been associated with defects in the IL11RA gene.
Competing interests: None declared
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Acknowledgments
We are grateful to:
Dr. Violetta Anastasiadou, Dr. Marianna Kousparou, and Dr. Andreas Hadjidemetriou, from Archbishop Makarios III Children’s Hospital in Nicosia, for offering their patients to be included in this study.
Dr. Kyproula Christodoulou, Head of the Department of Neurogenetics of the Cyprus Institute of Neurology & Genetics, for her help regarding haplotype analysis.
Mr. Mark Greenslade and Ms. Sarah Burton-Jones from Bristol Genetics Laboratory, Southmead Hospital, Bristol, for excellent technical support.
We would also like to thank all the children and their parents for participating in this study and for their cooperation.
This work was funded by Telethon Cyprus and by the Cyprus Research Promotion Foundation (Project PENEK/0609/64 was cofinanced by the European Regional Development Fund and the Republic of Cyprus through the Research Promotion Foundation).
Disclosure Statement: The authors declare no conflict of interest.
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Communicated by: Gerard T. Berry, MD
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Synopsis
A novel contiguous deletion eliminates all GALT exons as well as the non-translated sequences of the adjacent interleukin 11 receptor alpha (IL11RA) gene causing classic galactosemia with additional phenotypic abnormalities such as craniosynostosis.
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Rena Papachristoforou, Petros Petrou, Hilary Sawyer, Maggie Williams, and Anthi Drousiotou declare that they have no conflict of interest.
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All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in this study.
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Papachristoforou, R., Petrou, P.P., Sawyer, H., Williams, M., Drousiotou, A. (2013). A Novel Large Deletion Encompassing the Whole of the Galactose-1-Phosphate Uridyltransferase (GALT) Gene and Extending into the Adjacent Interleukin 11 Receptor Alpha (IL11RA) Gene Causes Classic Galactosemia Associated with Additional Phenotypic Abnormalities. In: Zschocke, J., Gibson, K., Brown, G., Morava, E., Peters, V. (eds) JIMD Reports - Volume 12. JIMD Reports, vol 12. Springer, Cham. https://doi.org/10.1007/8904_2013_249
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DOI: https://doi.org/10.1007/8904_2013_249
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