Purification and properties of the ribonucleic acid-dependent ribonucleic acid polymerase from Halobacterium cutirubrum

Biochem J. 1972 Jul;128(4):755-62. doi: 10.1042/bj1280755.

Abstract

1. The RNA-dependent RNA polymerase from Halobacterium cutirubrum was purified to electrophoretic homogeneity. 2. It requires a single-stranded molecule of RNA or polyribonucleotide as template. 3. Nearest-neighbour analyses of the products formed on random poly(A,U) or alternating poly(A-U) templates and base analysis of the product of synthesis directed by wheat-germ RNA prove that the template is copied accurately. 4. The enzyme initiates new chains with purine ribonucleoside triphosphates. 5. Sucrose-density-gradient analysis of the product indicates that it has a size distribution similar to that of the template. 6. Preliminary amino acid analysis of the RNA-dependent polymerase shows that it contains much less serine than either of the subunits of H. cutirubrum DNA-dependent RNA polymerase. 7. The RNA-dependent enzyme is unable to substitute for either subunit of the DNA-dependent polymerase, and both the latter are devoid of RNA-dependent activity.

MeSH terms

  • Amino Acids / analysis
  • Base Sequence
  • Cells, Cultured
  • Centrifugation, Density Gradient
  • DNA-Directed RNA Polymerases / analysis
  • DNA-Directed RNA Polymerases / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Halobacterium / enzymology*
  • Phosphorus Isotopes
  • RNA, Bacterial
  • Serine / analysis
  • Templates, Genetic

Substances

  • Amino Acids
  • Phosphorus Isotopes
  • RNA, Bacterial
  • Serine
  • DNA-Directed RNA Polymerases