synthesis The cellular processes of DNA replication and transcription involve DNA and RNA synthesis, respectively. DNA synthesis uses dNTPs as substrates, while RNA synthesis uses rNTPs as substrates. NTPs cannot be converted directly to dNTPs. DNA contains four different nitrogenous bases: adenine, guanine, cytosine and thymine. RNA also contains adenine, guanine, and cytosine, but replaces thymine with uracil. Thus, DNA synthesis requires dATP, dGTP, dCTP, and dTTP as substrates, while RNA synthesis requires ATP, GTP, CTP, and UTP. Nucleic acid synthesis is catalyzed by either DNA polymerase or RNA polymerase for DNA and RNA synthesis respectively. These enzymes covalently link the free -OH group on the 3’ carbon of a growing chain of nucleotides to the α-phosphate on the 5’ carbon of the next (d)NTP, releasing the β- and γ-phosphate groups as pyrophosphate (PPi). This results in a phosphodiester linkage between the two (d)NTPs. The release of PPi provides the energy necessary for the reaction to occur. It is important to note that nucleic acid synthesis occurs exclusively in the 5’ to 3’ direction. Nucleoside triphosphate metabolism Given their importance in the cell, the synthesis and degradation of nucleoside triphosphates is under tight control. This section focuses on nucleoside triphosphate metabolism in humans, but the process is fairly conserved among species. Nucleoside triphosphates cannot be absorbed well, so all nucleoside triphosphates are typically made de novo. The synthesis of ATP and GTP (purines) differs from the synthesis of CTP, TTP, and UTP (pyrimidines). Both purine and pyrimidine synthesis use phosphoribosyl pyrophosphate (PRPP) as a starting molecule. The conversion of NTPs to dNTPs can only be done in the diphosphate form. Typically a NTP has one phosphate removed to become a NDP, then is converted to a dNDP by an enzyme called ribonucleotide reductase, then a phosphate is added back to give a dNTP. Purine synthesis A nitrogenous base called hypoxanthine is assembled directly onto PRPP. This results in a nucleotide called inosine monophosphate (IMP). IMP is then converted to either a precursor to AMP or GMP. Once AMP or GMP are formed, they can be phosphorylated by ATP to their diphosphate and triphosphate forms. Purine synthesis is regulated by the allosteric inhibition of IMP formation by the adenine or guanine nucleotides. AMP and GMP also competitively inhibit the formation of their precursors from IMP. Pyrimidine synthesis A nitrogenous base called orotate is synthesized independently of PRPP. After orotate is made