Reverse transcriptase in archaebacteria. Purification and characterization of a primase-reverse-transcriptase complex from Halobacterium halobium.
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.
Ben-Mahrez
K
K
Institut Jacques Monod, Centre National de la Recherche Scientifique, Université Paris, France.
Sorokine
I
I
Nakayama
M
M
Kohiyama
M
M
D002845:Chromatography; D002852:Chromatography, Ion Exchange; D019915:DNA Primase; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D004722:Endoribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D006217:Halobacterium / Q000201:enzymology*; D006882:Hydroxyapatites; D007700:Kinetics; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification* / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000302:isolation & purification* / Q000378:metabolism; D016914:Ribonuclease H
D006882:Hydroxyapatites; D009097:Multienzyme Complexes; C067164:primase-reverse transcriptase complex; D017886:Durapatite; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D012194:RNA-Directed DNA Polymerase; D004259:DNA-Directed DNA Polymerase; D004722:Endoribonucleases; D016914:Ribonuclease H