Escherichia coli dGTP triphosphohydrolase is inhibited by gene 1.2 protein of bacteriophage T7.
Escherichia coli has a unique enzyme, deoxyguanosine triphosphate triphosphohydrolase (dGTPase) that cleaves dGTP into deoxyguanosine and tripolyphosphate. An E. coli mutant, optA1, has a 50-fold increased level of the dGTPase (Beauchamp, B.B., and Richardson, C.C. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 2563-2567). Successful infection of E. coli optA1 by bacteriophage T7 is dependent on a 10-kDa protein encoded by gene 1.2 of the phage. In this report we show that the gene 1.2 protein is a specific inhibitor of the E. coli dGTPase. Gene 1.2 protein inhibits dGTPase activity by forming a complex with the dGTPase with an apparent stoichiometry of two monomers of gene 1.2 protein/tetramer of dGTPase. The interaction is reversible with a half-life of the complex of 30 min and an apparent binding constant Ki of 35 nM. The binding of inhibitor of dGTPase is cooperative, indicating allosteric interactions between dGTPase subunits with a Hill coefficient of 1.7. The interaction is modulated differentially by DNA, RNA, and deoxyguanosine mono-, di-, and triphosphate. Both the binding of the substrate dGTP and of the inhibitor gene 1.2 protein induce conformational changes in dGTPase. The conformation of the enzyme in the presence of saturating concentrations of dGTP virtually prevents the association with, and the dissociation from, gene 1.2 protein.
Huber
H E
HE
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115.
Beauchamp
B B
BB
Richardson
C C
CC
D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013604:T-Phages; D014764:Viral Proteins / Q000378:metabolism / Q000494:pharmacology*
D003848:Deoxyguanine Nucleotides; D046911:Macromolecular Substances; D014764:Viral Proteins; D008345:Manganese; C029603:deoxyguanosine triphosphate; D004247:DNA; D010744:Phosphoric Monoester Hydrolases; D008274:Magnesium