Reverse transcriptase reads through a 2'-5'linkage and a 2'-thiophosphate in a template.
Avian myeloblastosis virus and Maloney murine leukemia virus RNase H-reverse transcriptases pause when they encounter a 2'-5' linkage or a 2'-thiophosphate in their template RNAs, but eventually read through these backbone modifications. Both reverse transcriptases pause after the 2'-5' linkage but before the 2'-thiophosphate. These results suggest that in the absence of precise information concerning the behavior of a given reverse transcriptase with respect to a particular lesion or modification, caution should be exercised in the interpretation of primer extension data that is being used to determine the existence of, or map the position of, a crosslink, site of chemical modification or non-standard linkage in an RNA template.
Lorsch
J R
JR
Department of Molecular Biology, Massachusetts General Hospital, Boston 02114, USA.
Bartel
D P
DP
Szostak
J W
JW
D009189:Avian Myeloblastosis Virus / Q000201:enzymology; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000201:enzymology; D010710:Phosphates / Q000737:chemistry*; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013698:Templates, Genetic
D010710:Phosphates; D012313:RNA; D004247:DNA; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D012194:RNA-Directed DNA Polymerase; C035638:thiophosphoric acid