Abstract
Discovery of DNA Polymerase
Highlights
DNA polymerase, an enzyme discovered in 1955, has the remarkable capacity to catalyze the template-directed synthesis of DNA [1, 2]
I will say at the outset that I am the author of this “Reflections” article, the discovery of DNA polymerase and the revelation that it is a template-directed enzyme resulted from the collective efforts of a small group that initially consisted of Maurice Bessman, Ernie Simms, and myself working with Arthur Kornberg in the Department of Microbiology at the Washington University School of Medicine
I had joined Arthur Kornberg’s laboratory in September of 1955 and had begun work on the purification of an enzyme in extracts of bacteriophage T2-infected cells that added a hydroxymethyl group to dCMP to form hydroxymethyl-dCMP. (In the T-even phages, cytosine is entirely replaced by hydroxymethylcytosine, which is present in various states of glucosylation [5, 6].) When Arthur showed me his results, I was tremendously excited by the possibility that they represented the first demonstration of DNA synthesis in vitro and I asked if I could put my project on hold and join him
Summary
I will say at the outset that I am the author of this “Reflections” article, the discovery of DNA polymerase and the revelation that it is a template-directed enzyme resulted from the collective efforts of a small group that initially consisted of Maurice Bessman, Ernie Simms, and myself working with Arthur Kornberg in the Department of Microbiology at the Washington University School of Medicine. Several months later Maurice Bessman arrived, and together with Ernie Simms, the four of us began to fractionate the activities responsible for the incorporation of the labeled thymidine into an acid-insoluble, DNase I-sensitive product. Once we had [32P]dTMP, we prepared ␣-[32P]dTTP by incubating our 32P-labeled dTMP with a partially purified nucleoside-diphosphate kinase and ATP and isolating the dTTP, which we carefully analyzed for thymidine, deoxyribose, and phosphate These were present in the ratio of 1:1:3. We began to fractionate the crude extract for dTTP incorporation into “DNA.”
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