Detection of RNA-dependent DNA polymerase activity in the Xiphophorus melanoma system.
DNA polymerases have been isolated from muscle and melanoma tissues of Xiphophorus, which are similar to retroviral RNA-dependent DNA polymerases as they prefer RNA to DNA templates. They appear to associate with submicroscopic structures which exhibit a density of about 1.13 g/ml after sucrose-density-gradient centrifugation. The RNA-dependent-DNA-polymerase-like enzymes could be separated from the DNA-dependent DNA polymerases by DEAE-cellulose chromatography. Further purification on phosphocellulose revealed that the muscle enzyme eluted at the void volume and at about 0.6 M KCl, whereas most of the melanoma enzyme eluted at 0.1 M KCl. Comparison of the template primer specificities of the muscle and melanoma enzymes with those of known DNA polymerases showed obvious similarities to the RNA-dependent DNA polymerase isolated from Rous sarcoma virus.
Genetisches Institut, Justus-Liebig-Universität, Giessen, Federal Republic of Germany.
W
W
Lüke
H
H
Petry
K
K
Hoefer
A
A
Anders
F
F
Anders
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000201:enzymology; D002499:Centrifugation, Density Gradient; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D005399:Fishes / Q000235:genetics; D008545:Melanoma / Q000201:enzymology*; D008854:Microscopy, Electron; D008861:Microsomes / Q000201:enzymology / Q000648:ultrastructure; D009132:Muscles / Q000201:enzymology; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification*; D013347:Subcellular Fractions / Q000201:enzymology
D012194:RNA-Directed DNA Polymerase