Purification and characterization of the DNA polymerase of human breast cancer particles.

Abstract

Previous studies have identified human breast tumor particles possessing many of the features characteristic of RNA tumor viruses. In addition to the expected size (600 S) and density (1.16 g/ml) these include possession of an outer membrane and an inner one surrounding a "core" containing a DNA polymerase and a large-molecular-weight (70S) RNA possessing detectable homology to the RNAs of the mouse mammary tumor virus (MMTV) and of the Mason-Pfizer monkey virus (MPMV). We report here the purification and characterization of the DNA polymerase from the human breast cancer particles. Its key properties are very similar to those ofthe RNA-dependent DNA nucleotidyltransferase (reverse transcriptase) found in MMTV and MPMV. Thus like these viral enzymes, the purified human breast cancer DNA polymerase exhibits the following three features that together distinguish the known viral reverse transcriptases from normal cellular DNA polymerases: (i) a strong preference for oligo(dT)-poly(rA) over oligo(dT)-poly(dA) as a template for the synthesis of poly(dT); (ii) the acceptance of the highly specific oligo(dG)-poly(rCm) as a template for the formation of poly(dG); (iii) the ability to use a viral RNA (AMV) as a template to fashion a faithful DNA complementary copy; and (iv) its preference for Mg++ over Mn++. In summary, the data described here on the enzyme of the human breast cancer particles add further evidence of similarities to the viral agents associated with the corresponding malignancies in the mouse and monkey models. To date, an enzyme with these properties has not been detected in normal breast tissues or in benign tumors of the breast.

• Authors: T Ohno, R W Sweet, D Dejak, S Spiegelman
• Journal: Proceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 74 Issue 2 Pg. 764-8 (Feb 1977) ISSN: 0027-8424 [Print] United States
• PMID: 265540 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Neoplasm
• Manganese
• DNA-Directed DNA Polymerase
• Magnesium

Topics

• Breast Neoplasms (enzymology)
• DNA, Neoplasm
• DNA-Directed DNA Polymerase (isolation & purification, metabolism)
• Enzyme Activation
• Female
• Humans
• Kinetics
• Magnesium (pharmacology)
• Manganese (pharmacology)
• Nucleic Acid Hybridization
• Subcellular Fractions (enzymology)
• Templates, Genetic