Reverse Transcriptase Pauses at N2-Methylguanine during in vitro Transcription of Escherichia coli 16S Ribosomal RNA
Abstract
A restriction fragment strand complementary to a sequence near the 3' end of Escherichia coli 16S rRNA has been used to prime reverse transcriptase (avian myeloblastosis virus RNA-directed DNA nucleotidyltransferase; deoxynucleosidetriphosphate:DNA deoxynucleotidyltransferase, EC 2.7.7.7). In addition to transcripts that were extended to the 5' end of the RNA, two major transcription intermediates were observed. These discrete-sized cDNA intermediates are the result of a kinetic barrier imposed by monomethylation of the amino group on guanine that participates in base-pairing. Both major transcription intermediates correspond to attenuation at the known positions of N2-methylguanine (m2G) in the rRNA sequence. The relaxation time for elongation of the cDNA through m2G is approximately 3 min. No other major kinetic pauses were observed in the 1340 bases transcribed.
- Publication:
-
Proceedings of the National Academy of Science
- Pub Date:
- August 1979
- DOI:
- 10.1073/pnas.76.8.3751
- Bibcode:
- 1979PNAS...76.3751Y