Mapping the tRNA Binding Site on the Surface of Human DNMT2 Methyltransferase

Publication type:
Journal article
Metadata:
Autoren
  • Tomasz P Jurkowski
  • Raghuvaran Shanmugam
  • Mark Helm
  • Albert Jeltsch
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000304783200007&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1021/bi3002659
Externe Identifier
  • Clarivate Analytics Document Solution ID: 952HL
  • PubMed Identifier: 22591353
ISSN
0006-2960
Ausgabe der Veröffentlichung
22
Zeitschrift
BIOCHEMISTRY
Paginierung
4438 - 4444
Datum der Veröffentlichung
2012
Status
Published
Titel
Mapping the tRNA Binding Site on the Surface of Human DNMT2 Methyltransferase
Sub types
  • Article
Ausgabe der Zeitschrift
51

Data source: Web of Science (Lite)

Other metadata sources:
Autoren
  • Tomasz P Jurkowski
  • Raghuvaran Shanmugam
  • Mark Helm
  • Albert Jeltsch
DOI
10.1021/bi3002659
eISSN
1520-4995
ISSN
0006-2960
Ausgabe der Veröffentlichung
22
Zeitschrift
Biochemistry
Sprache
en
Online publication date
2012
Paginierung
4438 - 4444
Datum der Veröffentlichung
2012
Status
Published
Herausgeber
American Chemical Society (ACS)
Herausgeber URL
http://dx.doi.org/10.1021/bi3002659
Datum der Datenerfassung
2023
Titel
Mapping the tRNA Binding Site on the Surface of Human DNMT2 Methyltransferase
Ausgabe der Zeitschrift
51

Data source: Crossref

Abstract
The DNMT2 enzyme methylates tRNA-Asp at position C38. Because there is no tRNA-Dnmt2 cocrystal structure available, we have mapped the tRNA binding site of DNMT2 by systematically mutating surface-exposed lysine and arginine residues to alanine and studying the tRNA methylation activity and binding of the corresponding variants. After mutating 20 lysine and arginine residues, we identified eight of them that caused large (>4-fold) decreases in catalytic activity. These residues cluster within and next to a surface cleft in the protein, which is large enough to accommodate the tRNA anticodon loop and stem. This cleft is located next to the binding pocket for the cofactor S-adenosyl-L-methionine, and the catalytic residues of DNMT2 are positioned at its walls or bottom. Many of the variants with strongly reduced catalytic activity showed only a weak loss of tRNA binding or even bound better to tRNA than wild-type DNMT2, which suggests that the enzyme induces some conformational changes in the tRNA in the transition state of the methyl group transfer reaction. Manual placement of tRNA into the structure suggests that DNMT2 mainly interacts with the anticodon stem and loop.
Addresses
  • Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Campus Ring 1, 28759 Bremen, Germany.
Autoren
  • Tomasz P Jurkowski
  • Raghuvaran Shanmugam
  • Mark Helm
  • Albert Jeltsch
DOI
10.1021/bi3002659
eISSN
1520-4995
Externe Identifier
  • PubMed Identifier: 22591353
Open access
false
ISSN
0006-2960
Ausgabe der Veröffentlichung
22
Zeitschrift
Biochemistry
Schlüsselwörter
  • Animals
  • Humans
  • Drosophila melanogaster
  • RNA, Transfer
  • Circular Dichroism
  • Cloning, Molecular
  • Amino Acid Substitution
  • Mutagenesis, Site-Directed
  • Sequence Alignment
  • Binding Sites
  • Amino Acid Sequence
  • Nucleic Acid Conformation
  • Protein Conformation
  • Methylation
  • Models, Molecular
  • Molecular Sequence Data
  • DNA (Cytosine-5-)-Methyltransferases
Sprache
eng
Medium
Print-Electronic
Online publication date
2012
Paginierung
4438 - 4444
Datum der Veröffentlichung
2012
Status
Published
Datum der Datenerfassung
2012
Titel
Mapping the tRNA binding site on the surface of human DNMT2 methyltransferase.
Sub types
  • Research Support, Non-U.S. Gov't
  • Journal Article
Ausgabe der Zeitschrift
51

Data source: Europe PubMed Central

Abstract
The DNMT2 enzyme methylates tRNA-Asp at position C38. Because there is no tRNA-Dnmt2 cocrystal structure available, we have mapped the tRNA binding site of DNMT2 by systematically mutating surface-exposed lysine and arginine residues to alanine and studying the tRNA methylation activity and binding of the corresponding variants. After mutating 20 lysine and arginine residues, we identified eight of them that caused large (>4-fold) decreases in catalytic activity. These residues cluster within and next to a surface cleft in the protein, which is large enough to accommodate the tRNA anticodon loop and stem. This cleft is located next to the binding pocket for the cofactor S-adenosyl-L-methionine, and the catalytic residues of DNMT2 are positioned at its walls or bottom. Many of the variants with strongly reduced catalytic activity showed only a weak loss of tRNA binding or even bound better to tRNA than wild-type DNMT2, which suggests that the enzyme induces some conformational changes in the tRNA in the transition state of the methyl group transfer reaction. Manual placement of tRNA into the structure suggests that DNMT2 mainly interacts with the anticodon stem and loop.
Autoren
  • Tomasz P Jurkowski
  • Raghuvaran Shanmugam
  • Mark Helm
  • Albert Jeltsch
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/22591353
DOI
10.1021/bi3002659
eISSN
1520-4995
Ausgabe der Veröffentlichung
22
Zeitschrift
Biochemistry
Schlüsselwörter
  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Binding Sites
  • Circular Dichroism
  • Cloning, Molecular
  • DNA (Cytosine-5-)-Methyltransferases
  • Drosophila melanogaster
  • Humans
  • Methylation
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Nucleic Acid Conformation
  • Protein Conformation
  • RNA, Transfer
  • Sequence Alignment
Sprache
eng
Country
United States
Paginierung
4438 - 4444
Datum der Veröffentlichung
2012
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2012
Titel
Mapping the tRNA binding site on the surface of human DNMT2 methyltransferase.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
51

Data source: PubMed

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