Other

Dataset Information

0

Anomalous reverse transcription through chemical modifications in polyadenosine stretches


ABSTRACT: Thermostable reverse transcriptases are workhorse enzymes underlying nearly all modern techniques for RNA structure mapping and for transcriptome-wide discovery of RNA chemical modifications. Despite their wide use, these enzymes’ behaviors at chemical modified nucleotides remain poorly understood. Wellington-Oguri et al. recently reported an apparent loss of chemical modification within putatively unstructured polyadenosine stretches modified by dimethyl sulfate or 2’ hydroxyl acylation, as probed by reverse transcription. Here, re-analysis of these and other publicly available data, capillary electrophoresis experiments on chemically modified RNAs, and nuclear magnetic resonance spectroscopy on A 12 and variants show that this effect is unlikely to arise from an unusual structure of polyadenosine. Instead, tests of different reverse transcriptases on chemically modified RNAs and molecules synthesized with single 1-methyladenosines implicate a previously uncharacterized reverse transcriptase behavior: near-quantitative bypass through chemical modifications within polyadenosine stretches. All tested natural and engineered reverse transcriptases (MMLV; SuperScript II, III, and IV; TGIRT-III; and MarathonRT) exhibit this anomalous bypass behavior. Accurate DMS-guided structure modeling of the polyadenylated HIV-1 3´ untranslated region RNA requires taking into account this anomaly. Our results suggest that poly(rA-dT) hybrid duplexes can trigger unexpectedly effective reverse transcriptase bypass and that chemical modifications in poly(A) mRNA tails may be generally undercounted.

ORGANISM(S): synthetic construct Human immunodeficiency virus

PROVIDER: GSE149061 | GEO | 2020/04/22

REPOSITORIES: GEO

Dataset's files

Source:
Action DRS
Other
Items per page:
1 - 1 of 1

Similar Datasets

| PRJNA627227 | ENA
2019-09-24 | GSE123365 | GEO
2017-09-26 | GSE97609 | GEO
2021-10-25 | GSE186464 | GEO
2023-06-01 | GSE174518 | GEO
2023-08-11 | GSE240200 | GEO
2022-08-10 | GSE209924 | GEO
2015-07-01 | GSE66550 | GEO
2024-08-08 | GSE269406 | GEO
2015-07-01 | E-GEOD-66550 | biostudies-arrayexpress