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Altered Pre-mRNA Splicing Caused by a Novel Intronic Mutation c.1443+5G>A in the Dihydropyrimidinase (DPYS) Gene.
Fujita Hlth Univ, Sch Med, Dept Pediat, Toyoake, Aichi 4701192, Japan.
Univ Amsterdam, Acad Med Ctr, Lab Genet Metab Dis, Meibergdreef 9, NL-1105 AZ Amsterdam, Netherlands.
MILS Int, Dept Res & Dev, Kanazawa, Ishikawa 9218105, Japan.
Capital Univ Med Sci, Beijing Childrens Hosp, Dept Neurol, Beijing 100045, Peoples R China.
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2016 (English)In: International Journal of Molecular Sciences, ISSN 1422-0067, E-ISSN 1422-0067, Vol. 17, no 1, 86Article in journal (Refereed) Published
Abstract [en]

Dihydropyrimidinase (DHP) deficiency is an autosomal recessive disease caused by mutations in the DPYS gene. Patients present with highly elevated levels of dihydrouracil and dihydrothymine in their urine, blood and cerebrospinal fluid. The analysis of the effect of mutations in DPYS on pre-mRNA splicing is hampered by the fact that DHP is primarily expressed in liver and kidney cells. The minigene approach can detect mRNA splicing aberrations using cells that do not express the endogenous mRNA. We have used a minigene-based approach to analyze the effects of a presumptive pre-mRNA splicing mutation in two newly identified Chinese pediatric patients with DHP deficiency. Mutation analysis of DPYS showed that both patients were compound heterozygous for a novel intronic mutation c.1443+5G>A in intron 8 and a previously described missense mutation c.1001A>G (p.Q334R) in exon 6. Wild-type and the mutated minigene constructs, containing exons 7, 8 and 9 of DPYS, yielded different splicing products after expression in HEK293 cells. The c.1443+5G>A mutation resulted in altered pre-mRNA splicing of the DPYS minigene construct with full skipping of exon 8. Analysis of the DHP crystal structure showed that the deletion of exon 8 severely affects folding, stability and homooligomerization of the enzyme as well as disruption of the catalytic site. Thus, the analysis suggests that the c.1443+5G>A mutation results in aberrant splicing of the pre-mRNA encoding DHP, underlying the DHP deficiency in two unrelated Chinese patients.

Place, publisher, year, edition, pages
2016. Vol. 17, no 1, 86
National Category
Medical Genetics
URN: urn:nbn:se:uu:diva-287443DOI: 10.3390/ijms17010086ISI: 000374583800082PubMedID: 26771602OAI: diva2:922739
Available from: 2016-04-25 Created: 2016-04-25 Last updated: 2016-07-07Bibliographically approved

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