Isolation and characterization of the gene coding for human cytidine deaminase

Demontis, Silvia; Terao, Mineko; Brivio, Massimo; Zanotta, Stefania; Bruschi, Maurizio; Garattini, Enrico

doi:10.1016/s0167-4781(98)00235-8

Cited by 31 publications

(20 citation statements)

References 31 publications

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“…7). Demontis et al (28) have reported that the CDA 5V-flanking region contains numerous potential transcription factor binding sites, so the present experiments represent merely an initial step in defining mechanisms responsible for the variations in transcription that we observed.…”

Section: Discussionmentioning

confidence: 73%

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

Gilbert¹,

Salavaggione²,

Ji³

et al. 2006

Clinical Cancer Research

150

128

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Purpose: Gemcitabine is a nucleoside analogue with activity against solid tumors. Gemcitabine metabolic inactivation is catalyzed by cytidine deaminase (CDA) or, after phosphorylation, by deoxycytidylate deaminase (DCTD). We set out to study the pharmacogenomics of CDA and DCTD. Experimental Design:The genes encoding CDA and DCTD were resequenced using DNA from 60 African American and 60 Caucasian American subjects. Expression constructs were created for nonsynonymous coding single nucleotide polymorphisms (cSNP) and reporter gene constructs were created for 5V -flanking region polymorphisms. Functional genomic studies were then conducted after the transfection of mammalian cells. Results: CDA resequencing revealed 17 polymorphisms, including one common nonsynonymous cSNP, 79 A>C (Lys27Gln). Recombinant Gln27 CDA had 66 F 5.1% (mean F SE) of the wild-type (WT) activity for gemcitabine but without a significant decrease in level of immunoreactive protein. The apparent K m (397 F 40 Amol/L) for the Gln27 allozyme was significantly higher than that for the WT (289 F 20 Amol/L; P < 0.025). CDA 5V-flanking region reporter gene studies showed significant differences among 5V -flanking region haplotypes in their ability to drive transcription. There were 29 SNPs in DCTD, including one nonsynonymous cSNP, 172 A>G (Asn58Asp), in Caucasian American DNA. Recombinant Asp58 DCTD had 11 F 1.4% of WT activity for gemcitabine monophosphate with a significantly elevated level of immunoreactive protein. No DCTD polymorphisms were observed in the initial 500 bp of the 5V -flanking region. Conclusions: These results suggest that pharmacogenomic variation in the deamination of gemcitabine and its monophosphate might contribute to variation in therapeutic response to this antineoplastic agent.

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Section: Discussionmentioning

confidence: 73%

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

Gilbert¹,

Salavaggione²,

Ji³

et al. 2006

Clinical Cancer Research

150

128

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“…Using 5Ј-RACE analysis of THP-1 cells, an unexpected finding was the identification of two different CDA transcripts-a long CDAlf transcript form, which has been previously reported (32), and a previously unreported short CDAsf transcript form that contained the complete sequences of exons 2, 3, and 4 plus up to 30 bp of intron 1 sequence. The presence of intron 1 sequences in the CDAsf 5Ј-UTRs implied that this form did not arise from alternate splicing but must be transcribed from its own promoter, presumably in intron 1.…”

Section: Discussionmentioning

confidence: 86%

The Role of Cytidine Deaminase and GATA1 Mutations in the Increased Cytosine Arabinoside Sensitivity of Down Syndrome Myeloblasts and Leukemia Cell Lines

Jensen

Stout

et al. 2004

Cancer Research

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“…The proximal promoter of the CDA gene contains binding sites for several transcription factors which regulate CDA expression in vivo and to have in vitro promoter activity (Watanabe and Uchida 1996;Demontis et al 1998;Ge et al 2004). Three of the identified SNPs, À92, À205 and À451, have minor allele frequencies greater than 0.15 and are informative for population studies.…”

Section: Resultsmentioning

confidence: 99%

“…Based on predicted changes in transcription factor binding sites and the ability to genotype by RFLP, three SNPs (-92A>G, -451C>T and -897C>A) were chosen for further investigation. These do not include the GATA1 sites proposed to be important in CDA regulation by Demontis et al (1998), nor the vitamin D response element speculated to be important by Watanabe and Uchida (1996) based on upregulation of CDA in cell lines by calcitrol. In a Dual Luciferase expression system, four out of five haplotypes containing these SNPs showed significantly higher relative luciferase expression compared with controls or with the lowest expression haplotype.…”

Section: Discussionmentioning

confidence: 99%

Identification of functional single nucleotide polymorphism haplotypes in the cytidine deaminase promoter

et al. 2006

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Cytidine deaminase (CDA) hydrolytically deaminates and irreversibly deactivates the chemotherapeutic agent cytosine arabinoside (Ara-C), a deoxycytidine analog used for treatment of acute leukemias and lymphomas. To determine if single nucleotide polymorphisms (SNPs) in the promoter region of CDA affected gene expression, we sequenced approximately 1.6 Kb upstream of the CDA translation initiation site and containing the proximal promoter of CDA. We identified 6 SNPs; -92A>G, -205C>G, -451C>T, -897C>A, -1075A>G and -1181G>A. Based on predicted changes in transcription factor binding sites, three SNPs (-92A>G, -451C>T and -897C>A) were chosen for further investigation. The five haplotypes segregating in the population were cloned into a luciferase expression plasmid, transfected into Cos-1 cells and reporter activity measured at 24 and 48 h. Four haplotypes showed an average expression which was 2.5-fold higher at 24 h (P<0.0001) and 3.3-fold higher at 48 h (P<0.0001) than the lowest expressing haplotype. When reanalyzed as single SNP genotypes, the differences in expression were significant, except for -897 C/A, at 24 h, but the magnitude of difference was reduced, suggesting that no single SNP completely accounts for the expression differences observed at the haplotype level. As predicted from the in vitro analysis, individuals homozygous for common haplotype (ACC/ACC) showed higher levels of CDA enzymatic activity as individuals heterozygous for the wild type and low expressing haplotype (ACC/ATC). As CDA promoter region haplotypes may influence Ara-C chemosensitivity, shown here in in vitro and in vivo studies, the clinical relevance of these findings should be examined.

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Isolation and characterization of the gene coding for human cytidine deaminase

Cited by 31 publications

References 31 publications

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

Gemcitabine Pharmacogenomics: Cytidine Deaminase and Deoxycytidylate Deaminase Gene Resequencing and Functional Genomics

The Role of Cytidine Deaminase and GATA1 Mutations in the Increased Cytosine Arabinoside Sensitivity of Down Syndrome Myeloblasts and Leukemia Cell Lines

Identification of functional single nucleotide polymorphism haplotypes in the cytidine deaminase promoter

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