Diastereoselectivity of 5-Methyluridine Osmylation Is Inverted inside an RNA Chain

Autoren

Lyudmil Tserovski
Mark Helm

DOI

10.1021/acs.bioconjchem.6b00403

eISSN

1520-4812

ISSN

1043-1802

Ausgabe der Veröffentlichung

9

Zeitschrift

Bioconjugate Chemistry

Sprache

en

Online publication date

2016

Paginierung

2188 - 2197

Datum der Veröffentlichung

2016

Status

Published

Herausgeber

American Chemical Society (ACS)

Herausgeber URL

http://dx.doi.org/10.1021/acs.bioconjchem.6b00403

Datum der Datenerfassung

2023

Titel

Diastereoselectivity of 5-Methyluridine Osmylation Is Inverted inside an RNA Chain

Ausgabe der Zeitschrift

27

Datenquelle: Crossref

Abstract

In this study, we investigated the reaction of the osmium tetroxide-bipyridine complex with pyrimidines in RNA. This reagent, which reacts with the diastereotopic 5-6 double bond, thus leading to the formation of two diastereomers, was used in the past to label thymidine and 5-methylcytosine in DNA. In light of the growing interest in post-transcriptional RNA modifications, we addressed the question of whether this reagent could be used for labeling of the naturally occurring RNA modifications 5-methylcytosine and 5-methyluridine. On nucleoside level, 5-methylcytosine and 5-methyluridine revealed a 5- and 12-fold preference, respectively, over their nonmethylated equivalents. Performing the reaction on an RNA level, we could show that the steric environment of a pentanucleotide has a major detrimental impact on the reaction rate of osmylation. Interestingly, this drop in reactivity was due to a dramatic change in diastereoselectivity, which in turn resulted from impediment of the preferred attack via the si side. Thus, while on the nucleoside level, the absolute configuration of the major product of osmylation of 5-methyluridine was (5R,6S)-5-methyluridine glycol-dioxoosmium-bipyridine, reaction with an RNA pentanucleotide afforded the corresponding (5S,6R)-diastereomer as the major product. The change in diastereoselectivity lead to an almost complete loss of selectivity toward 5-methylcytosine in a pentanucleotide context, while 5-methyluridine remained about 8 times more reactive than the canonical pyrimidines. On the basis of these findings, we evaluate the usefulness of osmium tetroxide-bipyridine as a potential label for the 5-methyluridine modification in transcriptome-wide studies.

Addresses

Institute of Pharmacy and Biochemistry, University of Mainz , D-55128 Mainz, Germany.

Autoren

Lyudmil Tserovski
Mark Helm

DOI

10.1021/acs.bioconjchem.6b00403

eISSN

1520-4812

Externe Identifier

PubMed Identifier: 27540864

Funding acknowledgements

Deutsche Forschungsgemeinschaft: HE3397/13-1
Deutsche Forschungsgemeinschaft: FOR 1082
Deutsche Forschungsgemeinschaft: HE 3397/6-2

Open access

false

ISSN

1043-1802

Ausgabe der Veröffentlichung

9

Zeitschrift

Bioconjugate chemistry

Schlüsselwörter

Organometallic Compounds
Pyridines
Pyrimidines
RNA
Uridine
Stereoisomerism

Sprache

eng

Medium

Print-Electronic

Online publication date

2016

Paginierung

2188 - 2197

Datum der Veröffentlichung

2016

Status

Published

Datum der Datenerfassung

2016

Titel

Diastereoselectivity of 5-Methyluridine Osmylation Is Inverted inside an RNA Chain.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

27

Datenquelle: Europe PubMed Central

Abstract

In this study, we investigated the reaction of the osmium tetroxide-bipyridine complex with pyrimidines in RNA. This reagent, which reacts with the diastereotopic 5-6 double bond, thus leading to the formation of two diastereomers, was used in the past to label thymidine and 5-methylcytosine in DNA. In light of the growing interest in post-transcriptional RNA modifications, we addressed the question of whether this reagent could be used for labeling of the naturally occurring RNA modifications 5-methylcytosine and 5-methyluridine. On nucleoside level, 5-methylcytosine and 5-methyluridine revealed a 5- and 12-fold preference, respectively, over their nonmethylated equivalents. Performing the reaction on an RNA level, we could show that the steric environment of a pentanucleotide has a major detrimental impact on the reaction rate of osmylation. Interestingly, this drop in reactivity was due to a dramatic change in diastereoselectivity, which in turn resulted from impediment of the preferred attack via the si side. Thus, while on the nucleoside level, the absolute configuration of the major product of osmylation of 5-methyluridine was (5R,6S)-5-methyluridine glycol-dioxoosmium-bipyridine, reaction with an RNA pentanucleotide afforded the corresponding (5S,6R)-diastereomer as the major product. The change in diastereoselectivity lead to an almost complete loss of selectivity toward 5-methylcytosine in a pentanucleotide context, while 5-methyluridine remained about 8 times more reactive than the canonical pyrimidines. On the basis of these findings, we evaluate the usefulness of osmium tetroxide-bipyridine as a potential label for the 5-methyluridine modification in transcriptome-wide studies.

Autoren

Lyudmil Tserovski
Mark Helm

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/27540864

DOI

10.1021/acs.bioconjchem.6b00403

eISSN

1520-4812

Ausgabe der Veröffentlichung

9

Zeitschrift

Bioconjug Chem

Schlüsselwörter

Organometallic Compounds
Pyridines
Pyrimidines
RNA
Stereoisomerism
Uridine

Sprache

eng

Country

United States

Paginierung

2188 - 2197

Datum der Veröffentlichung

2016

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2017

Titel

Diastereoselectivity of 5-Methyluridine Osmylation Is Inverted inside an RNA Chain.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

27

Datenquelle: PubMed

Diastereoselectivity of 5-Methyluridine Osmylation Is Inverted inside an RNA Chain

Werkzeuge