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Synthesis of Purine Ribonucleotides

Formation of deoxyribonucleotides
Ribonucleotide reductase catalyzes the synthesis of deoxyribonucleotide. The reductant is NADPH. Thioredoxin transfers electrons from NADPH for reduction of 2’-OH of ribose. dTMP is formed by thymidylate synthase by methylation of deoxy uridine monophosphate.

Synthesis of Pyrimidine Ribonucleotides

AB - A medium is described which can be used for the selection of somatic cell hybrids between HPRT- and APRT- mutant cells. This medium, called GAMA, contains as its relevant constituents the following: the purines guanine and adenine and the purine biosynthetic pathway inhibitors mycophenolic acid, which blocks conversion of adenine ribonucleotides to guanine ribonucleotides, and azaserine, which blocks de novo purine synthesis.

Protein Lounge: Synthesis of Pyrimidine Ribonucleotides

Synthesis of purine ribonucleotides
IMP is synthesized from ribose 5-phosphate. There are 11 reactions in the formation of IMP. IMP is converted to GMP and AMP with the help of ATP and GTP respectively. Nucleoside monophosphates are converted to nucleoside diphosphates by base specific monophosphate kinases. Purine nucleotide synthesis is regulated by feedback inhibitor – AMP, GMP and IMP. An important regulatory factor is the availability of PRPP. Salvage pathway for purines is observed in RBC and the brain. Free purines are salvaged by APRTase and HGPRTase enzymes

Synthesis of pyrimidine ribonucleotides
Pyrimidine ring is synthesized as free pyrimidine and then it is incorporated into the nucleotide. 6 reactions are involved in the synthesis of UMP. UDP and UTP are synthesized from UMP with the help of ATP. CTP is formed by adding an amino group from glutamine. Pyrimidine can also be salvaged using PRPP. In orotic aciduria, excretion of large amount of orotic acid is observed. It results from the deficiency of either orotate phospho ribosyl transferase or OMP decarboxylase.

N2 - A medium is described which can be used for the selection of somatic cell hybrids between HPRT- and APRT- mutant cells. This medium, called GAMA, contains as its relevant constituents the following: the purines guanine and adenine and the purine biosynthetic pathway inhibitors mycophenolic acid, which blocks conversion of adenine ribonucleotides to guanine ribonucleotides, and azaserine, which blocks de novo purine synthesis.

Purine and Pyrimidine Metabolism - EHSL

Figure 1 shows the time course of labeled RNA synthesis using 1 µg control template with Biotin-16-UTP and Fluorescein-12-UTP following the above reaction setup.

Modified ribonucleotides reduce transcription efficiency; therefore, lower transcription yields should be expected as compared to transcription using unmodified NTPs.

A medium is described which can be used for the selection of somatic cell hybrids between HPRT- and APRT- mutant cells. This medium, called GAMA, contains as its relevant constituents the following: the purines guanine and adenine and the purine biosynthetic pathway inhibitors mycophenolic acid, which blocks conversion of adenine ribonucleotides to guanine ribonucleotides, and azaserine, which blocks de novo purine synthesis.

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