Synthesis of signals for de novo DNA methylation in Neurospora crassa.
Identifieur interne : 003192 ( Main/Exploration ); précédent : 003191; suivant : 003193Synthesis of signals for de novo DNA methylation in Neurospora crassa.
Auteurs : Hisashi Tamaru [États-Unis] ; Eric U. SelkerSource :
- Molecular and cellular biology [ 0270-7306 ] ; 2003.
Descripteurs français
- KwdFr :
- ADN fongique (génétique), ADN fongique (métabolisme), ADN recombiné (physiologie), Appariement de bases (physiologie), Cytosine (métabolisme), Données de séquences moléculaires, Dosage biologique, Motifs AT-hook (physiologie), Mutation ponctuelle, Méthylation de l'ADN, Neurospora crassa, Protéine HMGB1 (métabolisme), Relation structure-activité, Séquence nucléotidique, Séquence riche en AT (physiologie), Séquences répétées d'acides nucléiques (physiologie), Test de retard de migration électrophorétique, Transduction du signal (physiologie).
- MESH :
- génétique : ADN fongique.
- métabolisme : ADN fongique, Cytosine, Protéine HMGB1.
- physiologie : ADN recombiné, Appariement de bases, Motifs AT-hook, Séquence riche en AT, Séquences répétées d'acides nucléiques, Transduction du signal.
- Données de séquences moléculaires, Dosage biologique, Mutation ponctuelle, Méthylation de l'ADN, Neurospora crassa, Relation structure-activité, Séquence nucléotidique, Test de retard de migration électrophorétique.
English descriptors
- KwdEn :
- AT Rich Sequence (physiology), AT-Hook Motifs (physiology), Base Pairing (physiology), Base Sequence, Biological Assay, Cytosine (metabolism), DNA Methylation, DNA, Fungal (genetics), DNA, Fungal (metabolism), DNA, Recombinant (physiology), Electrophoretic Mobility Shift Assay, HMGB1 Protein (metabolism), Molecular Sequence Data, Neurospora crassa, Point Mutation, Repetitive Sequences, Nucleic Acid (physiology), Signal Transduction (physiology), Structure-Activity Relationship.
- MESH :
- chemical , genetics : DNA, Fungal.
- chemical , metabolism : Cytosine, DNA, Fungal, HMGB1 Protein.
- physiology : AT Rich Sequence, AT-Hook Motifs, Base Pairing, DNA, Recombinant, Repetitive Sequences, Nucleic Acid, Signal Transduction.
- Base Sequence, Biological Assay, DNA Methylation, Electrophoretic Mobility Shift Assay, Molecular Sequence Data, Neurospora crassa, Point Mutation, Structure-Activity Relationship.
Abstract
Most 5-methylcytosine in Neurospora crassa occurs in A:T-rich sequences high in TpA dinucleotides, hallmarks of repeat-induced point mutation. To investigate how such sequences induce methylation, we developed a sensitive in vivo system. Tests of various 25- to 100-bp synthetic DNA sequences revealed that both T and A residues were required on a given strand to induce appreciable methylation. Segments composed of (TAAA)(n) or (TTAA)(n) were the most potent signals; 25-mers induced robust methylation at the special test site, and a 75-mer induced methylation elsewhere. G:C base pairs inhibited methylation, and cytosines 5' of ApT dinucleotides were particularly inhibitory. Weak signals could be strengthened by extending their lengths. A:T tracts as short as two were found to cooperate to induce methylation. Distamycin, which, like the AT-hook DNA binding motif found in proteins such as mammalian HMG-I, binds to the minor groove of A:T-rich sequences, suppressed DNA methylation and gene silencing. We also found a correlation between the strength of methylation signals and their binding to an AT-hook protein (HMG-I) and to activities in a Neurospora extract. We propose that de novo DNA methylation in Neurospora cells is triggered by cooperative recognition of the minor groove of multiple short A:T tracts. Similarities between sequences subjected to repeat-induced point mutation in Neurospora crassa and A:T-rich repeated sequences in heterochromatin in other organisms suggest that related mechanisms control silent chromatin in fungi, plants, and animals.
DOI: 10.1128/mcb.23.7.2379-2394.2003
PubMed: 12640122
Affiliations:
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Le document en format XML
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<term>Base Sequence</term>
<term>Biological Assay</term>
<term>Cytosine (metabolism)</term>
<term>DNA Methylation</term>
<term>DNA, Fungal (genetics)</term>
<term>DNA, Fungal (metabolism)</term>
<term>DNA, Recombinant (physiology)</term>
<term>Electrophoretic Mobility Shift Assay</term>
<term>HMGB1 Protein (metabolism)</term>
<term>Molecular Sequence Data</term>
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<term>Point Mutation</term>
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<term>Signal Transduction (physiology)</term>
<term>Structure-Activity Relationship</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>ADN fongique (génétique)</term>
<term>ADN fongique (métabolisme)</term>
<term>ADN recombiné (physiologie)</term>
<term>Appariement de bases (physiologie)</term>
<term>Cytosine (métabolisme)</term>
<term>Données de séquences moléculaires</term>
<term>Dosage biologique</term>
<term>Motifs AT-hook (physiologie)</term>
<term>Mutation ponctuelle</term>
<term>Méthylation de l'ADN</term>
<term>Neurospora crassa</term>
<term>Protéine HMGB1 (métabolisme)</term>
<term>Relation structure-activité</term>
<term>Séquence nucléotidique</term>
<term>Séquence riche en AT (physiologie)</term>
<term>Séquences répétées d'acides nucléiques (physiologie)</term>
<term>Test de retard de migration électrophorétique</term>
<term>Transduction du signal (physiologie)</term>
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<term>HMGB1 Protein</term>
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<term>Appariement de bases</term>
<term>Motifs AT-hook</term>
<term>Séquence riche en AT</term>
<term>Séquences répétées d'acides nucléiques</term>
<term>Transduction du signal</term>
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<term>AT-Hook Motifs</term>
<term>Base Pairing</term>
<term>DNA, Recombinant</term>
<term>Repetitive Sequences, Nucleic Acid</term>
<term>Signal Transduction</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term>
<term>Biological Assay</term>
<term>DNA Methylation</term>
<term>Electrophoretic Mobility Shift Assay</term>
<term>Molecular Sequence Data</term>
<term>Neurospora crassa</term>
<term>Point Mutation</term>
<term>Structure-Activity Relationship</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Données de séquences moléculaires</term>
<term>Dosage biologique</term>
<term>Mutation ponctuelle</term>
<term>Méthylation de l'ADN</term>
<term>Neurospora crassa</term>
<term>Relation structure-activité</term>
<term>Séquence nucléotidique</term>
<term>Test de retard de migration électrophorétique</term>
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<front><div type="abstract" xml:lang="en">Most 5-methylcytosine in Neurospora crassa occurs in A:T-rich sequences high in TpA dinucleotides, hallmarks of repeat-induced point mutation. To investigate how such sequences induce methylation, we developed a sensitive in vivo system. Tests of various 25- to 100-bp synthetic DNA sequences revealed that both T and A residues were required on a given strand to induce appreciable methylation. Segments composed of (TAAA)(n) or (TTAA)(n) were the most potent signals; 25-mers induced robust methylation at the special test site, and a 75-mer induced methylation elsewhere. G:C base pairs inhibited methylation, and cytosines 5' of ApT dinucleotides were particularly inhibitory. Weak signals could be strengthened by extending their lengths. A:T tracts as short as two were found to cooperate to induce methylation. Distamycin, which, like the AT-hook DNA binding motif found in proteins such as mammalian HMG-I, binds to the minor groove of A:T-rich sequences, suppressed DNA methylation and gene silencing. We also found a correlation between the strength of methylation signals and their binding to an AT-hook protein (HMG-I) and to activities in a Neurospora extract. We propose that de novo DNA methylation in Neurospora cells is triggered by cooperative recognition of the minor groove of multiple short A:T tracts. Similarities between sequences subjected to repeat-induced point mutation in Neurospora crassa and A:T-rich repeated sequences in heterochromatin in other organisms suggest that related mechanisms control silent chromatin in fungi, plants, and animals.</div>
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