Reverse transcriptase in archaebacteria. Purification and characterization of a primase-reverse-transcriptase complex from Halobacterium halobium

Eur J Biochem. 1991 Jan 1;195(1):157-62. doi: 10.1111/j.1432-1033.1991.tb15689.x.

Abstract

A primase-reverse-transcriptase of Halobacterium halobium was purified by column chromatography on DEAE-cellulose, hydroxyapatite and carboxymethyl-cellulose, followed by sedimentation on a glycerol gradient. The enzyme is a multifunctional enzyme containing reverse transcriptase. DNA polymerase and RNase H activities and does not require a performed primer to initiate DNA synthesis. Using a single-stranded DNA as template, this enzyme synthesizes oligonucleotides (8-12 bases) that can be used a primer by Escherichia coli DNA nucleotidyltransferase I (DNA polymerase I, Klenow fragment). Two polypeptides of 67 and 57 kDa were found after 14750-fold purification of the enzyme.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography
  • Chromatography, Ion Exchange
  • DNA Primase
  • DNA Replication
  • DNA-Directed DNA Polymerase / isolation & purification*
  • DNA-Directed DNA Polymerase / metabolism
  • Durapatite
  • Electrophoresis, Polyacrylamide Gel
  • Endoribonucleases / isolation & purification*
  • Endoribonucleases / metabolism
  • Halobacterium / enzymology*
  • Hydroxyapatites
  • Kinetics
  • Molecular Weight
  • Multienzyme Complexes / isolation & purification*
  • Multienzyme Complexes / metabolism
  • RNA Nucleotidyltransferases / isolation & purification*
  • RNA Nucleotidyltransferases / metabolism
  • RNA-Directed DNA Polymerase / isolation & purification*
  • RNA-Directed DNA Polymerase / metabolism
  • Ribonuclease H

Substances

  • Hydroxyapatites
  • Multienzyme Complexes
  • primase-reverse transcriptase complex
  • Durapatite
  • DNA Primase
  • RNA Nucleotidyltransferases
  • RNA-Directed DNA Polymerase
  • DNA-Directed DNA Polymerase
  • Endoribonucleases
  • Ribonuclease H