Re-examination by improved reverse transcriptase assay of DNA polymerase in platelets from myelodysplastic disease patients

Biochem Int. 1989 Nov;19(5):1133-41.

Abstract

A number of reports have suggested that platelets from polycythemia vera patients contain reverse transcriptase activity that might be correlated with C-type retrovirus-like particles. As described herein, we devised a new assay method for reverse transcriptase activity using MS-2 phage RNA hybridized with synthetic oligodeoxynucleotide (18-mer) as a template/primer. Using this new, sensitive assay method, we examined the platelet enzyme. By the conventional assay method using poly(rA)-oligo(dT), extracts of platelets showed a considerable amount of incorporation. However, by the new assay method using MS-2 RNA, no incorporation was observed. The poly(rA)-oligo(dT)-dependent activity was purified on Mono Q column, and it was shown that this activity coincided with that of DNA polymerase gamma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Platelets / enzymology*
  • Chromatography
  • DNA-Directed DNA Polymerase / analysis*
  • Humans
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / enzymology*
  • Magnesium / pharmacology
  • Manganese / physiology
  • Polycythemia Vera / enzymology*
  • RNA-Directed DNA Polymerase / metabolism*
  • RNA-Directed DNA Polymerase / physiology

Substances

  • Manganese
  • RNA-Directed DNA Polymerase
  • DNA-Directed DNA Polymerase
  • Magnesium