Abstract
Dihydropyrimidine dehydrogenase (DPD) is the initial and rate-limiting enzyme in the catabolism of the pyrimidine bases uracil and thymine, as well as of the widely used chemotherapeutic drug 5-fluorouracil (5FU). Analysis of the DPD gene (DPYD) in two patients presenting with complete DPD deficiency and the parents of an affected child showed the presence of three novel mutations, including one splice site mutation IVS11+1G→T and the missense mutations 731A→C (E244V) and 1651G→A (A551T). The G→T mutation in the invariant GT splice donor site flanking exon 11 (IVS11+1G→T) created a cryptic splice site within exon 11. As a consequence, a 141-bp fragment encoding the aminoacid residues 400–446 of the primary sequence of the DPD protein was missing in the mature DPD mRNA. Analysis of the crystal structure of pig DPD suggested that the E244V mutation might interfere with the electron flow between NADPH and the pyrimidine binding site of DPD. The A551T point mutation might prevent binding of the prosthetic group FMN and affect folding of the DPD protein. The identification of these novel mutations in DPYD will allow the identification of patients with an increased risk of developing severe 5FU-associated toxicity.
References
Au, K.M., Lai, C.K., Yuen, Y.P., Shek, C.C., Lam, C.W., and Chan, A.Y.W. (2003). Diagnosis of dihydropyrimidine dehydrogenase deficiency in a neonate with thymine-uraciluria. Hong Kong Med. J.9, 130–132.Search in Google Scholar
Collie-Duguid, E.S.R., Etienne, M.C., Milano, G.A., and McLeod, H.L. (2000). Known variant DPYD alleles do not explain DPD deficiency in cancer patients. Pharmacogenetics10, 217–223.10.1097/00008571-200004000-00002Search in Google Scholar
Diasio, R.B., Beavers, T.L., and Carpenter, J.T. (1988). Familial deficiency of dihydropyrimidine dehydrogenase. Biochemical basis for familial pyrimidinemia and severe 5-fluorouracil-induced toxicity. J. Clin. Invest.81, 47–51.Search in Google Scholar
Dobritzsch, D., Schneider, G., Schnackerz, K.D., and Lindqvist, Y. (2001). Crystal structure of dihydropyrimidine dehydrogenase, a major determinant of the pharmacokinetics of the anti-cancer drug 5-fluorouracil. EMBO J.20, 650–660.10.1093/emboj/20.4.650Search in Google Scholar
Enns, G.M., Barkovich, A.J., van Kuilenburg, A.B.P., Manning, M., Sanger, T., Witt, D.R., and van Gennip A.H. (2004). Head imaging abnormalities in dihydropyrimidine dehydrogenase deficiency. J. Inherit. Metab. Dis.27, 513–522.10.1023/B:BOLI.0000037350.24142.d5Search in Google Scholar
Esnouf, R.M. (1997). An extensively modified version of MolScript that includes greatly enhanced coloring capabilities. J. Mol. Graph. Model.15, 132–134.10.1016/S1093-3263(97)00021-1Search in Google Scholar
Fernandez-Salguero, P.M., Sapone, A., Wei, X., Holt, J.R., Jones, S., Idle, J.R., and Gonzales, F.J. (1997) Lack of correlation between phenotype and genotype for the polymorphically expressed dihydropyrimidine dehydrogenase in a family of Pakistani origin. Pharmacogenetics7, 161–163.10.1097/00008571-199704000-00012Search in Google Scholar
Gross, E., Ullrich, T., Seck, K., Mueller, V., de Wit, M., von Schilling C., Meindl, A., Schmitt, M., and Kiechle, M. (2003). Detailed analysis of five mutations in dihydropyrimidine dehydrogenase detected in cancer patients with 5-fluorouracil-related side effects. Hum. Mutat.22, 498.10.1002/humu.9201Search in Google Scholar
Johnson, M.R., Hageboutros, A., Wang, K., High, L., Smith, J.B., and Diasio, R.B. (1999). Life-threatening toxicity in a dihydropyrimidine dehydrogenase-deficient patient after treatment with topical 5-fluorouracil. Clin. Cancer Res.5, 2006–2011.Search in Google Scholar
Johnson, M.R., Wang, K., and Diasio, R.B. (2002). Profound dihydropyrimidine dehydrogenase deficiency resulting from a novel compound heterozygote genotype. Clin. Cancer Res.8, 768–774.Search in Google Scholar
Kraulis, P.J. (1991). MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallogr.24, 946–950.10.1107/S0021889891004399Search in Google Scholar
Mattison, L.K., Johnson, M.R., and Diasio, R.B. (2002). A comparative analysis of translated dihydropyrimidine dehydrogenase cDNA; conservation of functional domains and relevance to genetic polymorphisms. Pharmacogenetics12, 133–144.10.1097/00008571-200203000-00007Search in Google Scholar
Merritt, E.A. and Bacon, D.J. (1997). Raster 3D: photorealistic molecular graphics. Methods Enzymol.277, 505–524.10.1016/S0076-6879(97)77028-9Search in Google Scholar
Shapiro, M.B. and Senapathy, P. (1987). RNA splice junctions of different classes of eukaryotes: sequence statistics and functional implications in gene expression. Nucleic Acids Res.15, 7155–7174.10.1093/nar/15.17.7155Search in Google Scholar PubMed PubMed Central
van Gennip, A.H., Abeling, N.G.G.M., Vreken, P., and van Kuilenburg, A.B.P. (1997). Inborn errors of pyrimidine degradation: clinical, biochemical and molecular aspects. J. Inherit. Metab. Dis.20, 203–213.10.1023/A:1005356806329Search in Google Scholar
van Kuilenburg, A.B.P. (2004). Dihydropyrimidine dehydrogenase and the efficacy and toxicity of 5-fluorouracil. Eur. J. Cancer40, 939–950.10.1016/j.ejca.2003.12.004Search in Google Scholar PubMed
van Kuilenburg, A.B.P., Vreken, P., Abeling, N.G.G.M., Bakker, H.D., Meinsma, J.R., van Lenthe, H., De Abreu, R.A., Smeitink, J.A.M., Kayserili, H., Apak, M.Y., et al. (1999). Genotype and phenotype in patients with dihydropyrimidine dehydrogenase deficiency. Hum. Genet.104, 1–9.10.1007/PL00008711Search in Google Scholar PubMed
van Kuilenburg, A.B.P., Haasjes, J., Richel, D.J., Zoetekouw, L., van Lenthe, H., De Abreu, R.A., Maring, J.G., Vreken, P., and van Gennip, A.H. (2000a). Clinical implications of dihydropyrimidine dehydrogenase (DPD) deficiency in patients with severe 5-fluorouracil-associated toxicity: identification of new mutations in the DPD gene. Clin. Cancer Res.6, 4705–4712.Search in Google Scholar
van Kuilenburg, A.B.P., van Lenthe, H., Tromp, A., Veltman, P.C., and van Gennip A.H. (2000b). Pitfalls in the diagnosis of patients with a partial dihydropyrimidine dehydrogenase deficiency. Clin. Chem.46, 9–17.10.1093/clinchem/46.1.9Search in Google Scholar
van Kuilenburg, A.B.P., Dobritzsch, D., Meinsma, J.R., Haasjes, J., Waterham, H.R., Nowaczyk, M.J., Maropoulos, G.D., Hein, G., Kalhoff, H., Kirk, J.M. et al. (2002a). Novel disease-causing mutations in the dihydropyrimidine dehydrogenase gene interpreted by analysis of the three-dimensional protein structure. Biochem. J.364, 157–163.10.1042/bj3640157Search in Google Scholar PubMed PubMed Central
van Kuilenburg, A.B.P., Meinsma, R., Zoetekouw, L., and van Gennip, A.H. (2002b). Increased risk of grade IV neutropenia after administration of 5-fluorouracil due to a dihydropyrimidine dehydrogenase deficiency: high prevalence of the IVS14+1G→A mutation. Int. J. Cancer101, 253–258.10.1002/ijc.10599Search in Google Scholar PubMed
van Kuilenburg, A.B.P., Meinsma, J.R., Zonnenberg, B.A., Zoetekouw, L., Baas, F., Matsuda, K., Tamaki, N., and van Gennip, A.H. (2003). Dihydropyrimidinase deficiency and severe 5-fluorouracil toxicity. Clin. Cancer. Res.9, 4363–4367.Search in Google Scholar
van Kuilenburg, A.B.P., Stroomer, A.E.M., van Lenthe, H., Abeling, N.G.G.M., and van Gennip, A.H. (2004). New insights in dihydropyrimidine dehydrogenase deficiency: a pivotal role for β-aminoisobutyric acid? Biochem. J.379, 119–124.10.1042/bj20031463Search in Google Scholar PubMed PubMed Central
van Lenthe, H., van Kuilenburg, A.B.P., Ito, T., Bootsma, A.H., van Cruchten, A.G., Wada Y., and van Gennip, A.H. (2000). Defects in pyrimidine degradation identified by HPLC-electrospray tandem mass spectrometry of urine specimens or urine-soaked filter paper strips. Clin. Chem.46, 1916–1922.10.1093/clinchem/46.12.1916Search in Google Scholar
Vreken, P., van Kuilenburg, A.B.P., Meinsma, J.R., Smit, G.P.A., Bakker, H.D., De Abreu, R.A., and van Gennip, A.H. (1996). A point mutation in an invariant splice donor site leads to exon skipping in two unrelated Dutch patients with dihydropyrimidine dehydrogenase deficiency. J. Inherit. Metab. Dis.19, 645–654.10.1007/BF01799841Search in Google Scholar PubMed
Vreken, P., van Kuilenburg, A.B.P., Meinsma, J.R., and van Gennip A.H. (1997). Dihydropyrimidine dehydrogenase (DPD) deficiency: identification and expression of missense mutations C29R, R886H and R235W. Hum. Genet.101, 333–338.10.1007/s004390050637Search in Google Scholar PubMed
Wei, X., McLeod, H.L., McMurrough, J., and Gonzalez, F.J. (1996). Molecular basis of the human dihydropyrimidine dehydrogenase deficiency and 5-fluorouracil toxicity. J. Clin. Invest.98, 610–615.10.1172/JCI118830Search in Google Scholar PubMed PubMed Central
Wei, X., Elizondo, G., Sapone, A., McLeod, H.L., Raunio, H., and Gonzalez, F.J. (1998). Characterization of the human dihydropyrimidine dehydrogenase gene. Genomics51, 391–400.10.1006/geno.1998.5379Search in Google Scholar PubMed
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