Enzymatic Synthesis of Deoxyribonucleic Acid

XXI. UTILIZATION OF DEOXYRIBONUCLEOSIDE TRIPHOSPHATES BY ESCHERICHIA COLI CELLS

From the ¹ From the Department of Biochemistry, Stanford University School of Medicine, Palo Alto, California 94303

Escherichia coli cells incubated in tris(hydroxymethyl)-aminomethane buffer in the absence of divalent cations (Tris-ethylenediaminetetraacetate) or in Tris buffer in the presence of Mn⁺⁺ (Tris-Mn⁺⁺) utilize deoxyribonucleoside triphosphates for the intracellular synthesis of deoxyribonucleic acid. All four of the triphosphates are required, and they cannot be replaced by monophosphates or by ribonucleoside triphosphates. Based on the behavior of mutants deficient in endonuclease I and on other findings, the utilization of deoxynucleoside triphosphates after Tris-EDTA treatment is thought to be the result of (a) increased permeability and (b) the exposure of priming points for polymerase action on the cellular DNA. The specific effect of Mn⁺⁺ in the Tris-Mn⁺⁺ treatment is not explained; it does not depend on endonuclease I action nor does it, any more than Tris-EDTA, appear to involve the physiological operations of chromosomal replication.

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