A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine
DNA cytosine methylation (5-meC) is a widespread epigenetic mark associated to gene silencing. In plants, DEMETER-LIKE (DML) proteins typified by Arabidopsis REPRESSOR OF SILENCING 1 (ROS1) initiate active DNA demethylation by catalyzing 5-meC excision. DML proteins belong to the HhH-GPD superfamily...
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ftunivcordoba:oai:helvia.uco.es:10396/10870 2023-05-15T16:01:23+02:00 A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine Parrilla-Doblas, Jara Roldán-Arjona, Teresa Rodríguez Ariza, Rafael Ponferrada-Marín, María Isabel 2011 application/pdf http://hdl.handle.net/10396/10870 eng eng Oxford University Press http://hdl.handle.net/10396/10870 https://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess CC-BY-NC-ND Nucleic Acids Research 39 (4), 1473-1484 (2011) DNA Arabidopsis Enzymes info:eu-repo/semantics/article 2011 ftunivcordoba 2021-05-06T20:19:52Z DNA cytosine methylation (5-meC) is a widespread epigenetic mark associated to gene silencing. In plants, DEMETER-LIKE (DML) proteins typified by Arabidopsis REPRESSOR OF SILENCING 1 (ROS1) initiate active DNA demethylation by catalyzing 5-meC excision. DML proteins belong to the HhH-GPD superfamily, the largest and most functionally diverse group of DNA glycosylases, but the molecular properties that underlie their capacity to specifically recognize and excise 5-meC are largely unknown. We have found that sequence similarity to HhH-GPD enzymes in DML proteins is actually distributed over two non-contiguous segments connected by a predicted disordered region. We used homology-based modeling to locate candidate residues important for ROS1 function in both segments, and tested our predictions by site-specific mutagenesis. We found that amino acids T606 and D611 are essential for ROS1 DNA glycosylase activity, whereas mutations in either of two aromatic residues (F589 and Y1028) reverse the characteristic ROS1 preference for 5-meC over T. We also found evidence suggesting that ROS1 uses Q607 to flip out 5-meC, while the contiguous N608 residue contributes to sequence-context specificity. In addition to providing novel insights into the molecular basis of 5-meC excision, our results reveal that ROS1 and its DML homologs possess a discontinuous catalytic domain that is unprecedented among known DNA glycosylases. Article in Journal/Newspaper DML Helvia - Repositorio Institucional de la Universidad de Córdoba |
institution |
Open Polar |
collection |
Helvia - Repositorio Institucional de la Universidad de Córdoba |
op_collection_id |
ftunivcordoba |
language |
English |
topic |
DNA Arabidopsis Enzymes |
spellingShingle |
DNA Arabidopsis Enzymes Parrilla-Doblas, Jara Roldán-Arjona, Teresa Rodríguez Ariza, Rafael Ponferrada-Marín, María Isabel A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
topic_facet |
DNA Arabidopsis Enzymes |
description |
DNA cytosine methylation (5-meC) is a widespread epigenetic mark associated to gene silencing. In plants, DEMETER-LIKE (DML) proteins typified by Arabidopsis REPRESSOR OF SILENCING 1 (ROS1) initiate active DNA demethylation by catalyzing 5-meC excision. DML proteins belong to the HhH-GPD superfamily, the largest and most functionally diverse group of DNA glycosylases, but the molecular properties that underlie their capacity to specifically recognize and excise 5-meC are largely unknown. We have found that sequence similarity to HhH-GPD enzymes in DML proteins is actually distributed over two non-contiguous segments connected by a predicted disordered region. We used homology-based modeling to locate candidate residues important for ROS1 function in both segments, and tested our predictions by site-specific mutagenesis. We found that amino acids T606 and D611 are essential for ROS1 DNA glycosylase activity, whereas mutations in either of two aromatic residues (F589 and Y1028) reverse the characteristic ROS1 preference for 5-meC over T. We also found evidence suggesting that ROS1 uses Q607 to flip out 5-meC, while the contiguous N608 residue contributes to sequence-context specificity. In addition to providing novel insights into the molecular basis of 5-meC excision, our results reveal that ROS1 and its DML homologs possess a discontinuous catalytic domain that is unprecedented among known DNA glycosylases. |
format |
Article in Journal/Newspaper |
author |
Parrilla-Doblas, Jara Roldán-Arjona, Teresa Rodríguez Ariza, Rafael Ponferrada-Marín, María Isabel |
author_facet |
Parrilla-Doblas, Jara Roldán-Arjona, Teresa Rodríguez Ariza, Rafael Ponferrada-Marín, María Isabel |
author_sort |
Parrilla-Doblas, Jara |
title |
A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
title_short |
A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
title_full |
A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
title_fullStr |
A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
title_full_unstemmed |
A discontinuous DNA glycosylase domain in a family of enzymes that excise 5-methylcytosine |
title_sort |
discontinuous dna glycosylase domain in a family of enzymes that excise 5-methylcytosine |
publisher |
Oxford University Press |
publishDate |
2011 |
url |
http://hdl.handle.net/10396/10870 |
genre |
DML |
genre_facet |
DML |
op_source |
Nucleic Acids Research 39 (4), 1473-1484 (2011) |
op_relation |
http://hdl.handle.net/10396/10870 |
op_rights |
https://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
op_rightsnorm |
CC-BY-NC-ND |
_version_ |
1766397272908627968 |