Cargando…
Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
DNA N(6)-methyladenine modification plays an important role in regulating a variety of biological functions in bacteria. However, the mechanism of sequence-specific recognition in N(6)-methyladenine modification remains elusive. M1.HpyAVI, a DNA N(6)-adenine methyltransferase from Helicobacter pylor...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Impact Journals LLC
2016
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5173035/ https://www.ncbi.nlm.nih.gov/pubmed/27259995 http://dx.doi.org/10.18632/oncotarget.9692 |
_version_ | 1782484248313200640 |
---|---|
author | Ma, Bo Ma, Ji Liu, Dong Guo, Ling Chen, Huiling Ding, Jingjin Liu, Wei Zhang, Hongquan |
author_facet | Ma, Bo Ma, Ji Liu, Dong Guo, Ling Chen, Huiling Ding, Jingjin Liu, Wei Zhang, Hongquan |
author_sort | Ma, Bo |
collection | PubMed |
description | DNA N(6)-methyladenine modification plays an important role in regulating a variety of biological functions in bacteria. However, the mechanism of sequence-specific recognition in N(6)-methyladenine modification remains elusive. M1.HpyAVI, a DNA N(6)-adenine methyltransferase from Helicobacter pylori, shows more promiscuous substrate specificity than other enzymes. Here, we present the crystal structures of cofactor-free and AdoMet-bound structures of this enzyme, which were determined at resolutions of 3.0 Å and 3.1 Å, respectively. The core structure of M1.HpyAVI resembles the canonical AdoMet-dependent MTase fold, while the putative DNA binding regions considerably differ from those of the other MTases, which may account for the substrate promiscuity of this enzyme. Site-directed mutagenesis experiments identified residues D29 and E216 as crucial amino acids for cofactor binding and the methyl transfer activity of the enzyme, while P41, located in a highly flexible loop, playing a determinant role for substrate specificity. Taken together, our data revealed the structural basis underlying DNA N(6)-adenine methyltransferase substrate promiscuity. |
format | Online Article Text |
id | pubmed-5173035 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Impact Journals LLC |
record_format | MEDLINE/PubMed |
spelling | pubmed-51730352016-12-23 Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori Ma, Bo Ma, Ji Liu, Dong Guo, Ling Chen, Huiling Ding, Jingjin Liu, Wei Zhang, Hongquan Oncotarget Research Paper: Immunology DNA N(6)-methyladenine modification plays an important role in regulating a variety of biological functions in bacteria. However, the mechanism of sequence-specific recognition in N(6)-methyladenine modification remains elusive. M1.HpyAVI, a DNA N(6)-adenine methyltransferase from Helicobacter pylori, shows more promiscuous substrate specificity than other enzymes. Here, we present the crystal structures of cofactor-free and AdoMet-bound structures of this enzyme, which were determined at resolutions of 3.0 Å and 3.1 Å, respectively. The core structure of M1.HpyAVI resembles the canonical AdoMet-dependent MTase fold, while the putative DNA binding regions considerably differ from those of the other MTases, which may account for the substrate promiscuity of this enzyme. Site-directed mutagenesis experiments identified residues D29 and E216 as crucial amino acids for cofactor binding and the methyl transfer activity of the enzyme, while P41, located in a highly flexible loop, playing a determinant role for substrate specificity. Taken together, our data revealed the structural basis underlying DNA N(6)-adenine methyltransferase substrate promiscuity. Impact Journals LLC 2016-05-29 /pmc/articles/PMC5173035/ /pubmed/27259995 http://dx.doi.org/10.18632/oncotarget.9692 Text en Copyright: © 2016 Ma et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Paper: Immunology Ma, Bo Ma, Ji Liu, Dong Guo, Ling Chen, Huiling Ding, Jingjin Liu, Wei Zhang, Hongquan Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori |
title | Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
|
title_full | Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
|
title_fullStr | Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
|
title_full_unstemmed | Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
|
title_short | Biochemical and structural characterization of a DNA N(6)-adenine methyltransferase from Helicobacter pylori
|
title_sort | biochemical and structural characterization of a dna n(6)-adenine methyltransferase from helicobacter pylori |
topic | Research Paper: Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5173035/ https://www.ncbi.nlm.nih.gov/pubmed/27259995 http://dx.doi.org/10.18632/oncotarget.9692 |
work_keys_str_mv | AT mabo biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT maji biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT liudong biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT guoling biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT chenhuiling biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT dingjingjin biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT liuwei biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori AT zhanghongquan biochemicalandstructuralcharacterizationofadnan6adeninemethyltransferasefromhelicobacterpylori |