Allosteric control of the substrate specificity of the anaerobic ribonucleotide reductase from Escherichia coli.
The reduction of ribonucleotides is catalyzed by different enzymes in aerobic and anaerobic Escherichia coli, each with a different primary and quaternary structure. Here, we describe the allosteric regulation of the substrate specificity of the anaerobic ribonucleoside triphosphate reductase. The enzyme reduced ribonucleotides at a low basal rate. Reduction was stimulated up to 10-fold by an appropriate modulator (dGTP for ATP reduction, ATP for CTP and UTP reduction, and dTTP for GTP reduction). dGTP and dTTP inhibited the reduction of the "incorrect" substrate; dATP inhibited reduction of all four. From kinetic, effector binding, and competition experiments we conclude that the enzyme has two classes of sites, one that binds ATP and dATP and regulates pyrimidine ribonucleotide reduction ("pyrimidine site"), the other that binds dATP, dGTP, and dTTP and regulates purine ribonucleotide reduction ("purine site"). This model differs slightly from the model for the aerobic reductase, but the physiological consequences remain the same and explain how a single enzyme can provide a balanced supply of the four dNTPs. The similarity of a highly sophisticated control mechanism for the aerobic and anaerobic enzymes suggests that both arose by divergent evolution from a common ancestor, in spite of their different structures.
Eliasson
R
R
Department of Biochemistry 1, Medical Nobel Institute, Karolinska Institutet, Stockholm, Sweden.
Pontis
E
E
Sun
X
X
Reichard
P
P
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000494:Allosteric Regulation; D000693:Anaerobiosis; D001667:Binding, Competitive; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D012264:Ribonucleotide Reductases / Q000378:metabolism*; D013379:Substrate Specificity; D013942:Thymine Nucleotides / Q000378:metabolism
D003848:Deoxyguanine Nucleotides; D013942:Thymine Nucleotides; C029603:deoxyguanosine triphosphate; D000255:Adenosine Triphosphate; D012264:Ribonucleotide Reductases; C024157:thymidine 5'-triphosphate