<em>In vitro</em> tRNA Methylation Assay with the <em>Entamoeba histolytica</em> DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Ayala Tovy
- Benjamin Hofmann
- Mark Helm
- Serge Ankri
- DOI
- 10.3791/2390
- eISSN
- 1940-087X
- Ausgabe der Veröffentlichung
- 44
- Zeitschrift
- Journal of Visualized Experiments
- Sprache
- en
- Online publication date
- 2010
- Status
- Published online
- Herausgeber
- MyJove Corporation
- Herausgeber URL
- http://dx.doi.org/10.3791/2390
- Datum der Datenerfassung
- 2016
- Titel
- <em>In vitro</em> tRNA Methylation Assay with the <em>Entamoeba histolytica</em> DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
Datenquelle: Crossref
- Andere Metadatenquellen:
-
- Abstract
- Protozoan parasites are among the most devastating infectious agents of humans responsible for a variety of diseases including amebiasis, which is one of the three most common causes of death from parasitic disease. The agent of amebiasis is the amoeba parasite Entamoeba histolytica that exists under two stages: the infective cyst found in food or water and the invasive trophozoite living in the intestine. The clinical manifestations of amebiasis range from being asymptomatic to colitis, dysentery or liver abscesses. E. histolytica is one of the rare unicellular parasite with 5-methylcytosine (5mC) in its genome. It contains a single DNA methyltransferase, Ehmeth, that belongs to the Dnmt2 family. A role for Dnmt2 in the control of repetitive elements has been established in E. histolytica, Dictyostelium discoideum and Drosophila. Our recent work has shown that Ehmeth methylates tRNA(Asp), and this finding indicates that this enzyme has a dual DNA/tRNA(Asp) methyltransferase activity. This observation is in agreement with the dual activity that has been reported for D. discoideum and D. melanogaster. The functional significance of the DNA/tRNA specificity of Dnmt2 enzymes is still unknown. To address this question, a method to determine the tRNA methyltransferase activity of Dnmt2 proteins was established. In this video, we describe a straightforward approach to prepare an adequate tRNA substrate for Dnmt2 and a method to measure its tRNA methyltransferase activity.
- Addresses
- Faculty of Medicine, Rappaport Institute, Technion - Israel Institute of Technology.
- Autoren
- Ayala Tovy
- Benjamin Hofmann
- Mark Helm
- Serge Ankri
- DOI
- 10.3791/2390
- eISSN
- 1940-087X
- Externe Identifier
- PubMed Identifier: 21048666
- PubMed Central ID: PMC3185636
- Open access
- false
- ISSN
- 1940-087X
- Ausgabe der Veröffentlichung
- 44
- Zeitschrift
- Journal of visualized experiments : JoVE
- Schlüsselwörter
- Entamoeba histolytica
- Fungal Proteins
- RNA, Fungal
- RNA, Transfer
- Methylation
- DNA (Cytosine-5-)-Methyltransferases
- Sprache
- eng
- Medium
- Electronic
- Online publication date
- 2010
- Paginierung
- 2390
- Datum der Veröffentlichung
- 2010
- Status
- Published
- Datum der Datenerfassung
- 2010
- Titel
- In vitro tRNA methylation assay with the Entamoeba histolytica DNA and tRNA methyltransferase Dnmt2 (Ehmeth) enzyme.
- Sub types
- Video-Audio Media
- Research Support, Non-U.S. Gov't
- research-article
- Journal Article
Datenquelle: Europe PubMed Central
- Abstract
- Protozoan parasites are among the most devastating infectious agents of humans responsible for a variety of diseases including amebiasis, which is one of the three most common causes of death from parasitic disease. The agent of amebiasis is the amoeba parasite Entamoeba histolytica that exists under two stages: the infective cyst found in food or water and the invasive trophozoite living in the intestine. The clinical manifestations of amebiasis range from being asymptomatic to colitis, dysentery or liver abscesses. E. histolytica is one of the rare unicellular parasite with 5-methylcytosine (5mC) in its genome. It contains a single DNA methyltransferase, Ehmeth, that belongs to the Dnmt2 family. A role for Dnmt2 in the control of repetitive elements has been established in E. histolytica, Dictyostelium discoideum and Drosophila. Our recent work has shown that Ehmeth methylates tRNA(Asp), and this finding indicates that this enzyme has a dual DNA/tRNA(Asp) methyltransferase activity. This observation is in agreement with the dual activity that has been reported for D. discoideum and D. melanogaster. The functional significance of the DNA/tRNA specificity of Dnmt2 enzymes is still unknown. To address this question, a method to determine the tRNA methyltransferase activity of Dnmt2 proteins was established. In this video, we describe a straightforward approach to prepare an adequate tRNA substrate for Dnmt2 and a method to measure its tRNA methyltransferase activity.
- Autoren
- Ayala Tovy
- Benjamin Hofmann
- Mark Helm
- Serge Ankri
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/21048666
- DOI
- 10.3791/2390
- eISSN
- 1940-087X
- Externe Identifier
- PubMed Central ID: PMC3185636
- Ausgabe der Veröffentlichung
- 44
- Zeitschrift
- J Vis Exp
- Schlüsselwörter
- DNA (Cytosine-5-)-Methyltransferases
- Entamoeba histolytica
- Fungal Proteins
- Methylation
- RNA, Fungal
- RNA, Transfer
- Sprache
- eng
- Country
- United States
- PII
- 2390
- Datum der Veröffentlichung
- 2010
- Status
- Published online
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2010
- Titel
- In vitro tRNA methylation assay with the Entamoeba histolytica DNA and tRNA methyltransferase Dnmt2 (Ehmeth) enzyme.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Video-Audio Media
Datenquelle: PubMed
- Beziehungen:
- Eigentum von