Origin of pyrimidine deoxyribonucleotide pools in perfused rat heart: implications for 3'-azido-3'-deoxythymidine-dependent cardiotoxicity.
In adult non-replicating tissues such as heart, demand for dNTPs (deoxynucleoside triphosphates) is low but essential for mitochondrial DNA replication and nuclear DNA repair. dNTPs may be synthesized from salvage of deoxyribonucleosides or by reduction of ribonucleotides. We have hypothesized that the cardiac mitochondrial toxicity of the nucleoside analogue AZT (3'-azido-3'-deoxythymidine; known as zidovudine) is caused by inhibition of thymidine kinase 2 of the salvage pathway and subsequent TTP pool depletion. The extent to which this hypothesis has merit depends on how much the heart relies on thymidine phosphorylation for maintenance of the TTP pool. In the present study, we used isotopic tracing to demonstrate that both TTP and dCTP are solely synthesized by phosphorylation of thymidine and deoxycytidine respectively, with no evidence for synthesis from other precursors. We have also shown that UTP and CTP are synthesized by phosphorylation of uridine and cytidine respectively, with no detectable role for the de novo pyrimidine synthesis pathway. Lastly, we have demonstrated that AZT decreased the TTP pool by 50% in 30 min of perfusion, while having no effect on other dNTPs. In summary, the present study demonstrated that adult rat heart has a limited mechanism for dCTP and TTP synthesis and thus these pools may be more sensitive than replicating cells to drugs such as AZT that affect the salvage pathway.
Morris
Gerald W
GW
Indiana University School of Medicine South Bend, South Bend, IN 46617, U.S.A.
Iams
Tyler A
TA
Slepchenko
Kira G
KG
McKee
Edward E
EE
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003562:Cytidine / Q000378:metabolism; D003570:Cytidine Triphosphate / Q000378:metabolism; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D003841:Deoxycytidine / Q000378:metabolism; D003845:Deoxycytosine Nucleotides / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D003857:Deoxyuridine / Q000378:metabolism; D006321:Heart / Q000187:drug effects*; D066298:In Vitro Techniques; D008297:Male; D010477:Perfusion; D010766:Phosphorylation / Q000187:drug effects; D011742:Pyrimidine Nucleotides / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology* / Q000633:toxicity; D013942:Thymine Nucleotides / Q000378:metabolism; D014498:Uracil / Q000378:metabolism; D014529:Uridine / Q000378:metabolism; D014542:Uridine Monophosphate / Q000378:metabolism; D014544:Uridine Triphosphate / Q000378:metabolism; D015215:Zidovudine / Q000494:pharmacology* / Q000633:toxicity
D003838:Deoxyadenine Nucleotides; D003845:Deoxycytosine Nucleotides; D003848:Deoxyguanine Nucleotides; D011742:Pyrimidine Nucleotides; D018894:Reverse Transcriptase Inhibitors; D013942:Thymine Nucleotides; D003841:Deoxycytidine; C024107:2'-deoxycytidine 5'-triphosphate; D015215:Zidovudine; D014498:Uracil; D003562:Cytidine; D003570:Cytidine Triphosphate; D014542:Uridine Monophosphate; C024157:thymidine 5'-triphosphate; D014544:Uridine Triphosphate; D003857:Deoxyuridine; D014529:Uridine