Use of DNAzymes for site-specific analysis of ribonucleotide modifications
- Publication type:
- Journal article
- Metadata:
-
- Autoren
- Martin Hengesbach
- Madeleine Meusburger
- Frank Lyko
- Mark Helm
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000251698200018&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1261/rna.742708
- eISSN
- 1469-9001
- Externe Identifier
- Clarivate Analytics Document Solution ID: 242AZ
- PubMed Identifier: 17998290
- ISSN
- 1355-8382
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- RNA
- Schlüsselwörter
- DNAzyme
- tandem DNAzyme
- modification
- modified nucleotide
- modification enzymes
- Paginierung
- 180 - 187
- Datum der Veröffentlichung
- 2008
- Status
- Published
- Titel
- Use of DNAzymes for site-specific analysis of ribonucleotide modifications
- Sub types
- Article
- Ausgabe der Zeitschrift
- 14
Data source: Web of Science (Lite)
- Other metadata sources:
-
- Abstract
- <jats:p>Post-transcriptional ribonucleotide modifications are widespread and abundant processes that have not been analyzed adequately due to the lack of appropriate detection methods. Here, two methods for the analysis of modified nucleotides in RNA are presented that are based on the quantitative and site-specific DNAzyme-mediated cleavage of the target RNA at or near the site of modification. Quantitative RNA cleavage is achieved by cycling the DNAzyme and its RNA substrate through repeated periods of heating and cooling. In a first approach, DNAzyme-directed cleavage directly 5′ of the residue in question allows radioactive labeling of the newly freed 5′-OH. After complete enzymatic hydrolysis, the modification status can be assessed by two-dimensional thin layer chromatography. In a second approach, oligoribonucleotide fragments comprising the modification site are excised from the full-length RNA in an endonucleolytic fashion, using a tandem DNAzyme. The excised fragment is isolated by electrophoresis and submitted to further conventional analysis. These results establish DNAzymes as valuable tools for the site-specific and highly sensitive detection of ribonucleotide modifications.</jats:p>
- Autoren
- Martin Hengesbach
- Madeleine Meusburger
- Frank Lyko
- Mark Helm
- DOI
- 10.1261/rna.742708
- eISSN
- 1469-9001
- ISSN
- 1355-8382
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- RNA
- Sprache
- en
- Online publication date
- 2007
- Paginierung
- 180 - 187
- Datum der Veröffentlichung
- 2008
- Status
- Published
- Herausgeber
- Cold Spring Harbor Laboratory
- Herausgeber URL
- http://dx.doi.org/10.1261/rna.742708
- Datum der Datenerfassung
- 2021
- Titel
- Use of DNAzymes for site-specific analysis of ribonucleotide modifications
- Ausgabe der Zeitschrift
- 14
Data source: Crossref
- Abstract
- Post-transcriptional ribonucleotide modifications are widespread and abundant processes that have not been analyzed adequately due to the lack of appropriate detection methods. Here, two methods for the analysis of modified nucleotides in RNA are presented that are based on the quantitative and site-specific DNAzyme-mediated cleavage of the target RNA at or near the site of modification. Quantitative RNA cleavage is achieved by cycling the DNAzyme and its RNA substrate through repeated periods of heating and cooling. In a first approach, DNAzyme-directed cleavage directly 5' of the residue in question allows radioactive labeling of the newly freed 5'-OH. After complete enzymatic hydrolysis, the modification status can be assessed by two-dimensional thin layer chromatography. In a second approach, oligoribonucleotide fragments comprising the modification site are excised from the full-length RNA in an endonucleolytic fashion, using a tandem DNAzyme. The excised fragment is isolated by electrophoresis and submitted to further conventional analysis. These results establish DNAzymes as valuable tools for the site-specific and highly sensitive detection of ribonucleotide modifications.
- Addresses
- Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, 69120 Heidelberg, Germany.
- Autoren
- Martin Hengesbach
- Madeleine Meusburger
- Frank Lyko
- Mark Helm
- DOI
- 10.1261/rna.742708
- eISSN
- 1469-9001
- Externe Identifier
- PubMed Identifier: 17998290
- PubMed Central ID: PMC2151034
- Open access
- false
- ISSN
- 1355-8382
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- RNA (New York, N.Y.)
- Schlüsselwörter
- DNA, Catalytic
- RNA, Transfer, Lys
- DNA Primers
- Ribonucleotides
- Base Sequence
- Nucleic Acid Conformation
- Molecular Sequence Data
- Sprache
- eng
- Medium
- Print-Electronic
- Online publication date
- 2007
- Paginierung
- 180 - 187
- Datum der Veröffentlichung
- 2008
- Status
- Published
- Datum der Datenerfassung
- 2007
- Titel
- Use of DNAzymes for site-specific analysis of ribonucleotide modifications.
- Sub types
- Research Support, Non-U.S. Gov't
- research-article
- Journal Article
- Ausgabe der Zeitschrift
- 14
Files
http://rnajournal.cshlp.org/content/14/1/180.full.pdf https://europepmc.org/articles/PMC2151034?pdf=render
Data source: Europe PubMed Central
- Abstract
- Post-transcriptional ribonucleotide modifications are widespread and abundant processes that have not been analyzed adequately due to the lack of appropriate detection methods. Here, two methods for the analysis of modified nucleotides in RNA are presented that are based on the quantitative and site-specific DNAzyme-mediated cleavage of the target RNA at or near the site of modification. Quantitative RNA cleavage is achieved by cycling the DNAzyme and its RNA substrate through repeated periods of heating and cooling. In a first approach, DNAzyme-directed cleavage directly 5' of the residue in question allows radioactive labeling of the newly freed 5'-OH. After complete enzymatic hydrolysis, the modification status can be assessed by two-dimensional thin layer chromatography. In a second approach, oligoribonucleotide fragments comprising the modification site are excised from the full-length RNA in an endonucleolytic fashion, using a tandem DNAzyme. The excised fragment is isolated by electrophoresis and submitted to further conventional analysis. These results establish DNAzymes as valuable tools for the site-specific and highly sensitive detection of ribonucleotide modifications.
- Autoren
- Martin Hengesbach
- Madeleine Meusburger
- Frank Lyko
- Mark Helm
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/17998290
- DOI
- 10.1261/rna.742708
- eISSN
- 1469-9001
- Externe Identifier
- PubMed Central ID: PMC2151034
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- RNA
- Schlüsselwörter
- Base Sequence
- DNA Primers
- DNA, Catalytic
- Molecular Sequence Data
- Nucleic Acid Conformation
- RNA, Transfer, Lys
- Ribonucleotides
- Sprache
- eng
- Country
- United States
- Paginierung
- 180 - 187
- PII
- rna.742708
- Datum der Veröffentlichung
- 2008
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2008
- Titel
- Use of DNAzymes for site-specific analysis of ribonucleotide modifications.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 14
Data source: PubMed
- Beziehungen:
- Property of