Evidence for two forms of reverse transcriptase in human placenta of a patient with breast cancer. Purification and biochemical characterization of the enzymes.
Two DNA polymerases with properties of viral RNA-directed DNA polymerase were found in the placenta of a patient with breast cancer. Both enzyme activities were purified by column-chromatographic procedures or by preparative isoelectric focusing. The most distinguishing feature of the two enzymes is their specificity to transcribe (rA)n . (dT)12 or (rC)n . (dG)18. The two enzymes differ with respect to their elution profiles from the phosphocellulose column, isoelectric point, molecular weight, bivalent-cation requirements and thermal stability. Serological analysis of the (rA)n . (dT)12-activated enzyme showed that this enzyme is immunologically not related to DNA polymerase-gamma, or to any of the reverse transcriptases purified from retroviruses of avian, murine and subprimate origin. However, the activity of this enzyme was neutralized by antibodies to reverse transcriptase purified from human spleen of a patient with myelofibrosis [Chandra & Steel (1977) Biochem. J. 167, 513-524]. Attempts to purify reverse transcriptase of normal human placenta were repeatedly unsuccessful. Once the crude homogenate of normal placenta was freed from endogenous nucleic acids, no (rC)n . (dG)18-dependent activity cold be detected.U
D001943:Breast Neoplasms / Q000201:enzymology*; D002413:Cations, Divalent / Q000494:pharmacology; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D010920:Placenta / Q000201:enzymology*; D012367:RNA, Viral / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism
D002413:Cations, Divalent; D007527:Isoenzymes; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase