Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.4.2.8 (hypoxanthine-guanine phosphoribosyltransferase)
 2,527 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Cisplatin was reacted with a 184-base-pair sequence, exon 3, of human HPRT DNA in vitro. The binding sites were mapped by a primer extension method with T4 DNA polymerase and radioactive dCTP. Binding sites of cisplatin were indicated by the lengths of synthesized polynucleotides as determined by gel electrophoresis. Neighboring GG dinucleotides were highly preferred sites of binding by cisplatin, while less binding was noted to GXG, GA, AAA, and GXA. Analysis by densitometry revealed a 5-fold difference in binding among the GG sequences. The relative binding to a GGG sequence exceeded that of a GGGGGG sequence, suggesting that the number of Gs in a run did not determine the relative binding.
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PMID:Binding of cisplatin to specific sequences of human DNA in vitro. 318 85

The effects of introducing various DNA damage into pSV2gpt DNA on the subsequent expression of xanthine guanine phosphoribosyltransferase (XGPRT), after its transfection into two Walker 256 cell lines, one which is inherently sensitive only to difunctional agents while the other shows a normal sensitivity, have been examined. Both the sensitive (WS) and the relatively resistant (WR) cell lines were shown to be equally capable of both ligation of DNA double-strand breaks (although the efficiency varied with the actual site of the break) introduced into pSV2gpt and homologous recombination of pSV2gpt fragments (recombination events are thought to be important in the repair of DNA-DNA interstrand crosslinks). Reacting the plasmid with either the difunctional platinum compound, Cisplatin, or the monofunctional reacting Pt(Dien) caused a dose-dependent decrease in the subsequent expression of XGPRT. This decrease was about the same with either agent in either cell line when expressed as a function of dose of drug. However, when the actual binding of platinum to DNA by these compounds was measured, a large difference (due to the higher specific binding of Pt(Dien) to DNA) in the effects of the difunctional, as opposed to the monofunctional agent, was apparent and this was a reflection of the relative cytotoxicities of these compounds towards mammalian cells. Although at doses of Cisplatin equitoxic to WS and WR cells 20-fold less Pt is bound to the DNA of WS cells, no significant difference was seen on the expression of pSV2gpt, reacted with this agent, between WS or WR cells. Based upon a knowledge of the proportions of adducts formed in DNA reacted with Cisplatin, the lesion that inactivates expression of XGPRT was probably the intrastrand crosslink and it was calculated that due to the size of the plasmid, the interstrand crosslink was unlikely to be present at these inactivating doses. It is suggested that the inherent sensitivity of WS cells only to difunctional agents is due to their response to such relatively rare lesions such as a DNA-DNA interstrand crosslink.
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PMID:The effect of monofunctional or difunctional platinum adducts and of various other associated DNA damage on the expression of transfected DNA in mammalian cell lines sensitive or resistant to difunctional agents. 356 97

Previously, it has been shown that the V-H4 mutant of Chinese hamster V79 cells is homologous to Fanconi anemia (FA) group A cells. This hamster cell mutant shows a specific sensitivity to DNA cross-linking agents; therefore, the induction and repair of DNA cross-links were studied in V-H4 and wild-type V79 cells after cis-DDP treatment by the DNA alkaline elution technique. A significant difference in repair of these lesions in V-H4 and wild-type cells was observed. After the cis-DDP treatment (24 h) about 3 times more cross-links remained in V-H4 cells in comparison to the parental V79 cells. These results indicate that the process of cross-link repair in V-H4 cells is hampered when compared to that of wild-type cells. To assess the effect of slower removal of DNA cross-links on the mutability of V-H4, the induction of mutants at the hypoxanthine-guanine phosphoribosyltransferase locus (HPRT) by cis-DDP was studied in V-H4 and V79 cells. Despite the increased cytotoxicity of cis-DDP to V-H4 cells, the mutation induction at the HPRT locus was not significantly different in both cell lines, but when the frequency of the hprt mutants was plotted against survival, hypomutability was observed in V-H4 cells after the cis-DDP treatment.
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PMID:Mutagenic response and repair of cis-DDP-induced DNA cross-links in the Chinese hamster V79 cell mutant V-H4 which is homologous to Fanconi anemia (group A). 751 Mar 61

Oxaliplatin is a clinical anticancer drug with a pharmacological profile distinct from that of cisplatin. Our studies compared site- and region-specificity of lesions induced by oxaliplatin and cisplatin in naked and intracellular DNA, respectively. Oxaliplatin adducts in naked Simian virus 40 (SV40 DNA) were mapped by repetitive primer extension. The sites of oxaliplatin adducts were nearly identical to the sites of cisplatin adducts and were focused in G clusters and GNG motifs probably reflecting intrastrand cross-links. Although alkaline agarose electrophoresis of specific SV40 fragments showed that oxaliplatin formed interstrand cross-links, the levels of this lesion type were low. Drug-induced lesions in discrete loci of cellular DNA were assessed by the polymerase chain reaction stop assay in human tumor A2780 cells. Oxaliplatin at 200 microM induced approximately 1300, approximately 1500, approximately 800, and approximately 300 lesions/10(6) bp in the human beta-globin, c-myc, and HPRT genes and in mitochondrial DNA, respectively. Cisplatin formed two to six times more lesions in the same regions. For both drugs, lesion frequencies seem to parallel the density of drug-binding motifs in the nuclear regions, whereas mitochondrial DNA was disproportionately less affected. Despite less potent induction of DNA lesions, oxaliplatin was more cytotoxic than cisplatin against A2780 cells. Because our findings clearly demonstrate that oxaliplatin forms covalent adducts with a similar sequence- and region-specificity to that of cisplatin, other properties of oxaliplatin adducts, factors other than DNA binding, or both determine the unique features of the mechanism of action of oxaliplatin.
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PMID:Sequence- and region-specificity of oxaliplatin adducts in naked and cellular DNA. 980 12

The alkaline comet assay is a sensitive test for the detection of a variety of DNA lesions. However, crosslinks are not readily detected under standard test conditions. Recently, modifications have been introduced measuring crosslinks by determining the reduction of induced DNA migration. We used the comet assay to comparatively investigate in V79 cells the effect of three different crosslinkers: formaldehyde (FA), which predominantly induces DNA-protein crosslinks, cisplatin (DDP), which mainly produces DNA-DNA-intrastrand crosslinks, and mitomycin C (MMC), which mainly leads to DNA-DNA-interstrand crosslinks. In the standard alkaline comet assay, only MMC induced a slight increase in DNA migration at high toxic concentrations. FA and DDP did not induce any DNA migration under the test conditions used. In the modified comet assay, all three crosslinkers led to a clear reduction of gamma-ray-induced DNA migration. This reduction was seen in the case of FA parallel to the induction of cytotoxicity and SCE, while for MMC and DDP induction of cytotoxicity, SCE and HPRT gene mutations occurred at much lower concentrations than the effects in the comet assay. The DNA-DNA crosslinkers caused a reduction of induced DNA migration only at cytotoxic concentrations. Our results indicate that the modified comet assay protocol is a sensitive test for the detection of DNA-protein crosslinks. However, the results for MMC and DDP suggest that the modified protocol is not well suited for the evaluation of DNA-DNA crosslinkers. Furthermore, the relationship between crosslinking and genotoxicity seems to be very different for the three different types of crosslinking substances.
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PMID:Detection of crosslinks with the comet assay in relationship to genotoxicity and cytotoxicity. 1021 71