Characterization of the AIDS-associated retrovirus reverse transcriptase and optimal conditions for its detection in virions

Virology. 1985 Dec;147(2):326-35. doi: 10.1016/0042-6822(85)90135-7.

Abstract

The RNA-dependent DNA polymerase of the AIDS-associated retrovirus (ARV) gives highest activity with the synthetic template, poly(rA)oligo(dT) and prefers Mg2+ over Mn2+ as a divalent cation. It can use other template-primer combinations but with poly(rCm)oligo(dG), it prefers Mn2+ over Mg2+. Detection of ARV reverse transcriptase in culture fluids is substantially increased with an optimal KCl concentration and a special combination of EGTA and reducing agents. Our results indicate some distinguishing characteristics of the ARV reverse transcriptase and optimal conditions for its detection.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Cells, Cultured
  • Deltaretrovirus / enzymology*
  • Deltaretrovirus / growth & development
  • Egtazic Acid / pharmacology
  • Humans
  • Hydrogen-Ion Concentration
  • Magnesium / pharmacology
  • Manganese / pharmacology
  • Monocytes / enzymology
  • Monocytes / microbiology
  • Octoxynol
  • Oligodeoxyribonucleotides / metabolism
  • Poly A / metabolism
  • Polyethylene Glycols / pharmacology
  • Potassium Chloride / pharmacology
  • RNA-Directed DNA Polymerase / analysis
  • RNA-Directed DNA Polymerase / isolation & purification
  • RNA-Directed DNA Polymerase / metabolism*
  • Substrate Specificity
  • Sulfhydryl Reagents / pharmacology
  • Templates, Genetic
  • Virion / enzymology

Substances

  • Oligodeoxyribonucleotides
  • Sulfhydryl Reagents
  • Poly A
  • Polyethylene Glycols
  • Manganese
  • Egtazic Acid
  • poly(rA).oligo(dT)
  • Potassium Chloride
  • Octoxynol
  • RNA-Directed DNA Polymerase
  • Magnesium