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
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Aoki K, Wada Y (1988) Outcome of the patients detected by newborn screening in Japan. Acta Paediatr Jpn 30(4):429–434
Barbouth D, Slepak T, Klapper H, Lai K, Elsas LJ (2006) Prevention of a molecular misdiagnosis in galactosemia. Genet Med 8(3):178–182
Berry GT, Leslie N, Reynolds R, Yager CT, Segal S (2001) Evidence for alternate galactose oxidation in a patient with deletion of the galactose-1-phosphate uridyltransferase gene. Mol Genet Metab 72(4):316–321
Calderon FR, Phansalkar AR, Crockett DK, Miller M, Mao R (2007) Mutation database for the galactose-1-phosphate uridyltransferase (GALT) gene. Hum Mutat 28(10):939–943
Coffee B, Hjelm LN, DeLorenzo A, Courtney EM, Yu C, Muralidharan K (2006) Characterization of an unusual deletion of the galactose-1-phosphate uridyl transferase (GALT) gene. Genet Med 8(10):635–640
Coussens AK, Hughes IP, Wilkinson CR et al (2008) Identification of genes differentially expressed by prematurely fused human sutures using a novel in vivo – in vitro approach. Differentiation 76(5):531–545
Flach JE, Reichardt JK, Elsas LJ 2nd (1990) Sequence of a cDNA encoding human galactose-1-phosphate uridyl transferase. Mol Biol Med 7(4):365–369
Greber-Platzer S, Guldberg P, Scheibenreiter S et al (1997) Molecular heterogeneity of classical and Duarte galactosemia: mutation analysis by denaturing gradient gel electrophoresis. Hum Mutat 10(1):49–57
Holton JB, Walter JH, Tyfield LA (2001) Galactosemia. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited diseases, 8th edn. McGraw-Hill, New York, pp 1553–1587
Kalckar HM, Anderson EP, Isselbacher KJ (1956) Galactosemia, a congenital defect in a nucleotide transferase. Biochim Biophys Acta 20(1):262–268
Kong X, Matise TC (2005) MAP-O-MAT: internet-based linkage mapping. Bioinformatics 21(4):557–559
Lai K, Langley SD, Singh RH, Dembure PP, Hjelm LN, Elsas LJ 2nd (1996) A prevalent mutation for galactosemia among black Americans. J Pediatr 128(1):89–95
Leslie ND, Immerman EB, Flach JE, Florez M, Fridovich-Keil JL, Elsas LJ (1992) The human galactose-1-phosphate uridyltransferase gene. Genomics 14(2):474–480
Levy HL, Hammersen G (1978) Newborn screening for galactosemia and other galactose metabolic defects. J Pediatr 92(6):871–877
Li Y, Ptolemy AS, Harmonay L, Kellogg M, Berry GT (2010) Quantification of galactose-1-phosphate uridyltransferase enzyme activity by liquid chromatography-tandem mass spectrometry. Clin Chem 56(5):772–780
McVey M, Lee SE (2008) MMEJ repair of double-strand breaks (director’s cut): deleted sequences and alternative endings. Trends Genet 24(11):529–538
Murphy M, McHugh B, Tighe O et al (1999) Genetic basis of transferase-deficient galactosaemia in Ireland and the population history of the Irish Travellers. Eur J Hum Genet 7(5):549–554
Nieminen P, Morgan NV, Fenwick AL et al (2011) Inactivation of IL11 signaling causes craniosynostosis, delayed tooth eruption, and supernumerary teeth. Am J Hum Genet 89(1):67–81
Reichardt JK, Berg P (1988) Cloning and characterization of a cDNA encoding human galactose-1-phosphate uridyl transferase. Mol Biol Med 5(2):107–122
Schulpis K, Papakonstantinou ED, Michelakakis H, Podskarbi T, Patsouras A, Shin Y (1997) Screening for galactosaemia in Greece. Paediatr Perinat Epidemiol 11(4):436–440
Shih LY, Suslak L, Rosin I, Searle BM, Desposito F (1984) Gene dosage studies supporting localization of the structural gene for galactose-1-phosphate uridyl transferase (GALT) to band p13 of chromosome 9. Am J Med Genet 19(3):539–543
Tyfield L, Reichardt J, Fridovich-Keil J et al (1999) Classical galactosemia and mutations at the galactose-1-phosphate uridyl transferase (GALT) gene. Hum Mutat 13(6):417–430
Zekanowski C, Radomyska B, Bal J (1999) Molecular characterization of Polish patients with classical galactosaemia. J Inherit Metab Dis 22(5):679–682
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Additional information
Communicated by: Gerard T. Berry, MD
Electronic Supplementary Material
Below is the link to the electronic supplementary material.
Appendices
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.
Compliance with Ethics Guidelines
Conflict of Interest
Rena Papachristoforou, Petros Petrou, Hilary Sawyer, Maggie Williams, and Anthi Drousiotou declare that they have no conflict of interest.
Informed Consent
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.
Rights and permissions
Copyright information
© 2013 SSIEM and Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
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
Download citation
DOI: https://doi.org/10.1007/8904_2013_249
Received:
Revised:
Accepted:
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-03460-7
Online ISBN: 978-3-319-03461-4
eBook Packages: MedicineMedicine (R0)