B : dUMP Is Methylated to Form Thymine
The dTMP component of DNA synthesized by methylation of dUMP The d TMP is generated through the hydrolysis of dUTP by dUTP diphosphohydrolase (dUTPase):
dUTP +H2O -----dUMP +PP,
dTMP, once it is formed, is phosphorylated to form dTTP The apparent reason for the energetically wasteful process of dephosphorylating dUTP and rephosphorylating d TMP is that cells must minimize their concentra- tion of dUTP in order to prevent incorporation of uracil into their DNA. (the enzyme system that synthesizes DNA from dNTPs does not effi- ciently discriminate between dUTP and dTTP; Section 25-2A).
Human dUTPase is a homotrimer of 141-residue subunits. Its X-ray structure, determined by John Tainer, reveals the basis for the enzyme's exquisite specificity for dUTP Each subunit binds dUTP in a snug-fitting cavity that sterically excludes thymine's C5 methyl group via the side chains of conserved residues (Fig. 23-14a). The enzyme differentiates uracil from the similarly shaped cytosine via a set of hydrogen bonds that in part mimic adenine's base pairing interactions (Fig. 23-14b). The 2'-OH group of ri bose is likewise sterically excluded by the side chain of a conserved Tyr.