High processivity of the reverse transcriptase from a non-long terminal repeat retrotransposon.
R2 is a retrotransposable element that specifically inserts into the 28 S rRNA genes of arthropods. The element encodes a single protein with endonuclease activity that cleaves the 28 S gene target site and reverse transcriptase (RT) activity that uses the cleaved DNA to prime reverse transcription. Here we compare various properties of the R2 RT activity with those of the well characterized retroviral RT, avian myeloblastosis virus (AMV). In processivity assays using heterogeneous RNA templates, R2 RT can synthesize cDNA over twice the length of that synthesized by AMV RT and can synthesize cDNA over 4 times longer than AMV RT in assays with poly(rA) templates. The higher processivity of R2 RT compared with retroviral RTs is a result of the slower rate of dissociation of the enzyme from RNA templates. The elongation rates of the two enzymes are similar. Finally, a highly distinct property of the R2 RT, compared with retroviral enzymes, is its ability to displace RNA strands annealed to RNA templates during cDNA synthesis. We suggest that both the higher processivity and displacement properties of R2 RT compared with retroviral RT result from the greater affinity of the R2 protein for the RNA template upstream of its active site.
Bibillo
Arkadiusz
A
Department of Biology, University of Rochester, Rochester, New York 14627-0211, USA.
Eickbush
Thomas H
TH
D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D001483:Base Sequence; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D018626:Retroelements; D013698:Templates, Genetic
D012367:RNA, Viral; D018626:Retroelements; D012194:RNA-Directed DNA Polymerase