Using hetero-11-mers composed of wild type and mutant subunits to study tryptophan binding to TRAP and its role in activating RNA binding.
Identifieur interne : 003305 ( Main/Exploration ); précédent : 003304; suivant : 003306Using hetero-11-mers composed of wild type and mutant subunits to study tryptophan binding to TRAP and its role in activating RNA binding.
Auteurs : Pan T X. Li [États-Unis] ; Paul GollnickSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 2002.
Descripteurs français
- KwdFr :
- ARN bactérien (métabolisme), Bacillus subtilis (génétique), Biopolymères, Facteurs de transcription (), Facteurs de transcription (métabolisme), Liaison aux protéines, Modèles moléculaires, Protéines bactériennes (), Protéines bactériennes (métabolisme), Protéines de liaison à l'ARN (), Protéines de liaison à l'ARN (métabolisme), Tryptophane (métabolisme).
- MESH :
English descriptors
- KwdEn :
- Bacillus subtilis (genetics), Bacterial Proteins (chemistry), Bacterial Proteins (metabolism), Biopolymers, Models, Molecular, Protein Binding, RNA, Bacterial (metabolism), RNA-Binding Proteins (chemistry), RNA-Binding Proteins (metabolism), Transcription Factors (chemistry), Transcription Factors (metabolism), Tryptophan (metabolism).
- MESH :
- chemical , chemistry : Bacterial Proteins, RNA-Binding Proteins, Transcription Factors.
- genetics : Bacillus subtilis.
- chemical , metabolism : Bacterial Proteins, RNA, Bacterial, RNA-Binding Proteins, Transcription Factors, Tryptophan.
- chemical : Biopolymers, Models, Molecular, Protein Binding.
Abstract
Expression of genes involved in tryptophan metabolism in Bacillus subtilis is regulated by the TRAP protein in response to changes in l-tryptophan levels. TRAP binding to several RNA targets that contain between 9 and 11 (G/U)AG repeats regulates transcription and/or translation of these genes. TRAP consists of 11 identical subunits and is activated to bind RNA by binding up to 11 molecules of tryptophan. To investigate the mechanism by which tryptophan binding activates TRAP, we generated hetero-11-mers containing different proportions of subunits from wild type (WT) TRAP that bind tryptophan and from a mutant TRAP (Thr(25) to Ala) defective in tryptophan binding. Studies of these hetero-11-mers show that tryptophan-binding sites created from active subunits bind tryptophan with similar affinity to those in WT homo-11-mers, whereas sites containing the T25A substitution do not bind tryptophan. Hetero-11-mers with very few (one or two) bound tryptophans show only 10-fold lower affinity than WT TRAP for an RNA with 11 GAG repeats, whereas TRAP with no bound tryptophan shows no detectable binding to this RNA. We also demonstrate that tryptophan binding induces a conformational change in TRAP in the vicinity of the RNA-binding site, suggesting a possible mechanism for activation of RNA binding.
DOI: 10.1074/jbc.M205910200
PubMed: 12133840
Affiliations:
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Le document en format XML
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Li</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biological Sciences, State University of New York, Buffalo, New York 14260, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biological Sciences, State University of New York, Buffalo, New York 14260</wicri:regionArea><wicri:noRegion>New York 14260</wicri:noRegion></affiliation></author><author><name sortKey="Gollnick, Paul" sort="Gollnick, Paul" uniqKey="Gollnick P" first="Paul" last="Gollnick">Paul Gollnick</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2002">2002</date><idno type="RBID">pubmed:12133840</idno><idno type="pmid">12133840</idno><idno type="doi">10.1074/jbc.M205910200</idno><idno type="wicri:Area/PubMed/Corpus">002498</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002498</idno><idno type="wicri:Area/PubMed/Curation">002498</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002498</idno><idno type="wicri:Area/PubMed/Checkpoint">002343</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002343</idno><idno type="wicri:Area/Ncbi/Merge">000148</idno><idno type="wicri:Area/Ncbi/Curation">000148</idno><idno type="wicri:Area/Ncbi/Checkpoint">000148</idno><idno type="wicri:doubleKey">0021-9258:2002:Li P:using:hetero:mers</idno><idno type="wicri:Area/Main/Merge">003340</idno><idno type="wicri:Area/Main/Curation">003305</idno><idno type="wicri:Area/Main/Exploration">003305</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Using hetero-11-mers composed of wild type and mutant subunits to study tryptophan binding to TRAP and its role in activating RNA binding.</title><author><name sortKey="Li, Pan T X" sort="Li, Pan T X" uniqKey="Li P" first="Pan T X" last="Li">Pan T X. Li</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biological Sciences, State University of New York, Buffalo, New York 14260, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biological Sciences, State University of New York, Buffalo, New York 14260</wicri:regionArea><wicri:noRegion>New York 14260</wicri:noRegion></affiliation></author><author><name sortKey="Gollnick, Paul" sort="Gollnick, Paul" uniqKey="Gollnick P" first="Paul" last="Gollnick">Paul Gollnick</name></author></analytic><series><title level="j">The Journal of biological chemistry</title><idno type="ISSN">0021-9258</idno><imprint><date when="2002" type="published">2002</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Bacillus subtilis (genetics)</term><term>Bacterial Proteins (chemistry)</term><term>Bacterial Proteins (metabolism)</term><term>Biopolymers</term><term>Models, Molecular</term><term>Protein Binding</term><term>RNA, Bacterial (metabolism)</term><term>RNA-Binding Proteins (chemistry)</term><term>RNA-Binding Proteins (metabolism)</term><term>Transcription Factors (chemistry)</term><term>Transcription Factors (metabolism)</term><term>Tryptophan (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN bactérien (métabolisme)</term><term>Bacillus subtilis (génétique)</term><term>Biopolymères</term><term>Facteurs de transcription ()</term><term>Facteurs de transcription (métabolisme)</term><term>Liaison aux protéines</term><term>Modèles moléculaires</term><term>Protéines bactériennes ()</term><term>Protéines bactériennes (métabolisme)</term><term>Protéines de liaison à l'ARN ()</term><term>Protéines de liaison à l'ARN (métabolisme)</term><term>Tryptophane (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Bacterial Proteins</term><term>RNA-Binding Proteins</term><term>Transcription Factors</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Bacillus subtilis</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Bacillus subtilis</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Bacterial Proteins</term><term>RNA, Bacterial</term><term>RNA-Binding Proteins</term><term>Transcription Factors</term><term>Tryptophan</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN bactérien</term><term>Facteurs de transcription</term><term>Protéines bactériennes</term><term>Protéines de liaison à l'ARN</term><term>Tryptophane</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Biopolymers</term><term>Models, Molecular</term><term>Protein Binding</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Biopolymères</term><term>Facteurs de transcription</term><term>Liaison aux protéines</term><term>Modèles moléculaires</term><term>Protéines bactériennes</term><term>Protéines de liaison à l'ARN</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Expression of genes involved in tryptophan metabolism in Bacillus subtilis is regulated by the TRAP protein in response to changes in l-tryptophan levels. TRAP binding to several RNA targets that contain between 9 and 11 (G/U)AG repeats regulates transcription and/or translation of these genes. TRAP consists of 11 identical subunits and is activated to bind RNA by binding up to 11 molecules of tryptophan. To investigate the mechanism by which tryptophan binding activates TRAP, we generated hetero-11-mers containing different proportions of subunits from wild type (WT) TRAP that bind tryptophan and from a mutant TRAP (Thr(25) to Ala) defective in tryptophan binding. Studies of these hetero-11-mers show that tryptophan-binding sites created from active subunits bind tryptophan with similar affinity to those in WT homo-11-mers, whereas sites containing the T25A substitution do not bind tryptophan. Hetero-11-mers with very few (one or two) bound tryptophans show only 10-fold lower affinity than WT TRAP for an RNA with 11 GAG repeats, whereas TRAP with no bound tryptophan shows no detectable binding to this RNA. We also demonstrate that tryptophan binding induces a conformational change in TRAP in the vicinity of the RNA-binding site, suggesting a possible mechanism for activation of RNA binding.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Gollnick, Paul" sort="Gollnick, Paul" uniqKey="Gollnick P" first="Paul" last="Gollnick">Paul Gollnick</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Li, Pan T X" sort="Li, Pan T X" uniqKey="Li P" first="Pan T X" last="Li">Pan T X. Li</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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