RNA-dependent DNA polymerase from a cell line derived from the bone marrow of a patient with polycythemia vera.
A RNA-dependent DNA polymerase was isolated from a human cell line derived from the bone marrow of a patient with polycythemia vera. The purification procedure included chromatography on phosphocellulose and oligo(dT)-cellulose, and glycerol gradient centrifugation. The enzyme could be distinguished from polymerase A by salt elution from phosphocellulose, utilization of poly(rC) - oligo(dG) and its molecular size of about 70000, as determined by centrifugation. Throughout the purification procedure ribonuclease H activity was co-purified. Upon dodecylsulfate-polyacrylamide electrophoresis on microgradient gels two main bands with molecular weights of 68000 and 66000 and three minor bands were detected. The enzyme preferentially used poly(rA) - oligo(dT) as template-primer compared with poly(dA) - oligo(dT). It incorporated dGMP into polymer on poly(rC) - oligo(dG).
D001853:Bone Marrow / Q000201:enzymology*; D001854:Bone Marrow Cells; D002460:Cell Line; D002846:Chromatography, Affinity; D006801:Humans; D011087:Polycythemia Vera / Q000201:enzymology*; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D013698:Templates, Genetic
D012194:RNA-Directed DNA Polymerase