Re-examination by improved reverse transcriptase assay of DNA polymerase in platelets from myelodysplastic disease patients.
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.
Yoshida
S
S
Laboratory of Cancer Cell Biology, Nagoya University School of Medicine, Japan.
Yamada
H
H
Sakurai
T
T
Suzuki
M
M
Kojima
K
K
D001792:Blood Platelets / Q000201:enzymology*; D002845:Chromatography; D004259:DNA-Directed DNA Polymerase / Q000032:analysis*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000502:physiology; D011087:Polycythemia Vera / Q000201:enzymology*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism* / Q000502:physiology
D008345:Manganese; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase; D008274:Magnesium