Molecular basis of pyrimidine 5′-nucleotidase deficiency caused by 3 newly identified missense mutations (c.187T&gt;C, c.469G&gt;C and c.740T&gt;C) and a tabulation of known mutations

Chiarelli, Laurent R.; Morera, Simone M.; Galizzi, Alessandro; Fermo, Elisa; Zanella, Alberto; Valentini, Giovanna

doi:10.1016/j.bcmd.2007.10.005

Cited by 10 publications

(4 citation statements)

References 21 publications

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“…NT5C3 deficiency has been associated with hemolytic anemia [18, 26, 27], and a series of mutations were identified in patients with NT5C3 deficiency [15, 28, 29]. However, none of those mutations was observed in our samples.…”

Section: Discussionmentioning

confidence: 67%

Cytosolic 5′-nucleotidase III (NT5C3): gene sequence variation and functional genomics

Aksoy¹,

Zhu²,

Kalari³

et al. 2009

Pharmacogenetics and Genomics

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Background 5’-Nucleotidases play a critical role in nucleotide pool balance and in the metabolism of nucleoside analogs such as gemcitabine and cytosine arabinoside (AraC). We previously performed an expression array association study with gemcitabine and AraC cytotoxicity using 197 human lymphoblastoid cell lines. One gene that was significantly associated with gemcitabine cytotoxicity was a nucleotidase family member, NT5C3. Very little is known with regard to the pharmacogenomics of this family of enzymes. Methods We set out to identify common genetic variation in NT5C3 by resequencing the gene and to determine the effect of that variation on NT5C3 protein function and potential effect on response to cytidine analogues. We identified 61 NT5C3 polymorphisms, 48 of which were novel, by resequencing 240 ethnically defined DNA samples. Functional studies were performed with one nonsynonymous (G847C, Asp283His) and 4 synonymous cSNPs (T9C, C276T, T306C, and G759A), as well as three combined variants (T276/His283, T276/C306, T276/C9). Results The His283 and T276/His283 constructs showed decreased levels of enzyme activity and protein. Substrate kinetic analysis showed no significant differences in Km values between WT and His283 when CMP, AraCMP and GemMP were used as substrates. An association study between SNPs and NT5C3 expression in the 240 cell lines from which DNA was extracted to resequence NT5C3 identified 4 SNPs that were significantly associated with NT5C3 expression. EMSAs showed that two of those SNPs, I4(-114) and I6(9), altered DNA-protein binding patterns. These findings suggest that genetic variation in NT5C3 might affect protein function and potentially influence drug response.

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Section: Discussionmentioning

confidence: 67%

Cytosolic 5′-nucleotidase III (NT5C3): gene sequence variation and functional genomics

Aksoy¹,

Zhu²,

Kalari³

et al. 2009

Pharmacogenetics and Genomics

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“…An advantage of our findings is that the measurement of TKT expression levels by immunodetection or targeted MS would ease the process of diagnosing P5N deficiency in HA patients. This may serve as a preliminary screening and can be performed prior to the more laborious molecular characterization approaches [12,[46][47][48].…”

Section: Discussionmentioning

confidence: 99%

Proteomics reveals reduced expression of transketolase in pyrimidine 5′‐nucleotidase deficient patients

Barasa

Oirschot

Bianchi

et al. 2016

Proteomics Clinical Apps

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“…In contrast to erythrocyte membranopathies, osmotic fragility is ordinarily not increased [ 21 ]. Despite the fact that no known natural mutation is directly involved in the catalytic mechanism, a significant reduction in enzyme activity is observed in patients with P5ND [ 12 , 13 , 15 , 22 , 23 ]. The exact mechanisms leading to erythrocyte destruction and hemolytic anemia are still unclear, but it is known that pyrimidine nucleotide accumulation affects erythrocyte metabolism, including the decrease in intracellular pH, lower G6PD activity, and reduced ribose-phosphate pyrophosphokinase activity [ 12 , 15 , 24 – 26 ].…”

Section: Introductionmentioning

confidence: 99%

A rare mutation (p.F149del) of the NT5C3A gene is associated with pyrimidine 5′-nucleotidase deficiency

et al. 2022

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Pyrimidine 5′-nucleotidase deficiency is a rare erythrocyte enzymopathy. Here we report two cases of hemolytic anemia in brothers of Polish origin that are associated with a very rare mutation. Heterozygous deletion in the NT5C3A gene (c.444_446delGTT), inherited most likely from their asymptomatic mother, resulted in a single amino acid residue deletion (p.F149del) in cytosolic pyrimidine 5′-nucleotidase. However, only the mutated transcript was present in the reticulocyte transcriptome of both patients. Only residual activity of pyrimidine 5′-nucleotidase in the brothers’ erythrocytes could be observed when compared with the controls, including their asymptomatic father and sister. Western blot showed no sign of the presence of 5′-nucleotidase protein in the erythrocytes of both studied patients. The 2.5-fold reduction of the purine/pyrimidine ratio observed only in the brothers’ erythrocytes confirms the correlation of the results of molecular analysis, including whole-exome sequencing, with the phenotype of the pyrimidine 5′-nucleotidase deficiency. Altogether, our results may substantiate the hypothesis of the heterogeneity of the molecular basis of the defect involving both the mutation presented here and negative regulation of expression of the “normal” allele.

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Molecular basis of pyrimidine 5′-nucleotidase deficiency caused by 3 newly identified missense mutations (c.187T>C, c.469G>C and c.740T>C) and a tabulation of known mutations

Cited by 10 publications

References 21 publications

Cytosolic 5′-nucleotidase III (NT5C3): gene sequence variation and functional genomics

Cytosolic 5′-nucleotidase III (NT5C3): gene sequence variation and functional genomics

Proteomics reveals reduced expression of transketolase in pyrimidine 5′‐nucleotidase deficient patients

A rare mutation (p.F149del) of the NT5C3A gene is associated with pyrimidine 5′-nucleotidase deficiency

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