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Characterization of an Endogenous RNA-Dependent DNA Polymerase Associated with Murine Intracisternal A Particles

¹ Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20014

ABSTRACT

An RNA-dependent DNA polymerase associated with intracisternal A particles has been characterized. The enzyme required Mg²⁺ or Mn²⁺, dithiothreitol and the presence of all four deoxyribonucleoside triphosphates for the expression of maximal activity. Sensitivity of the endogenous RNA-dependent DNA polymerase activity to a low concentration of pancreatic ribonuclease in the presence of a high concentration of NaCl suggested that the enzyme might be utilizing the A particle endogenous RNA as template. Evidence in support of this was provided by analyses of early and late DNA products of the endogenous reaction by Cs₂SO₄ isopycnic gradient centrifugation and hybridization of purified 60 to 70S and 35S RNAs of A particles with the purified DNA product.