Creating hetero-11-mers composed of wild-type and mutant subunits to study RNA binding to TRAP.
Identifieur interne : 002375 ( PubMed/Checkpoint ); précédent : 002374; suivant : 002376Creating hetero-11-mers composed of wild-type and mutant subunits to study RNA binding to TRAP.
Auteurs : Pan T X. Li [États-Unis] ; David J. Scott ; Paul GollnickSource :
- The Journal of biological chemistry [ 0021-9258 ] ; 2002.
Descripteurs français
- KwdFr :
- ARN (métabolisme), Bacillus subtilis (métabolisme), Cinétique, Conformation des protéines, Dichroïsme circulaire, Dénaturation des protéines, Escherichia coli (métabolisme), Facteurs de transcription (), Facteurs temps, Guanidine (pharmacologie), Liaison aux protéines, Modèles chimiques, Mutation, Pliage des protéines, Protéines bactériennes, Protéines de liaison à l'ARN (), Relation dose-effet des médicaments, Structure secondaire des protéines, Tryptophane (), Ultracentrifugation.
- MESH :
- métabolisme : ARN, Bacillus subtilis, Escherichia coli.
- pharmacologie : Guanidine.
- Cinétique, Conformation des protéines, Dichroïsme circulaire, Dénaturation des protéines, Facteurs de transcription, Facteurs temps, Liaison aux protéines, Modèles chimiques, Mutation, Pliage des protéines, Protéines bactériennes, Protéines de liaison à l'ARN, Relation dose-effet des médicaments, Structure secondaire des protéines, Tryptophane, Ultracentrifugation.
English descriptors
- KwdEn :
- Bacillus subtilis (metabolism), Bacterial Proteins, Circular Dichroism, Dose-Response Relationship, Drug, Escherichia coli (metabolism), Guanidine (pharmacology), Kinetics, Models, Chemical, Mutation, Protein Binding, Protein Conformation, Protein Denaturation, Protein Folding, Protein Structure, Secondary, RNA (metabolism), RNA-Binding Proteins (chemistry), Time Factors, Transcription Factors (chemistry), Tryptophan (chemistry), Ultracentrifugation.
- MESH :
- chemical , chemistry : RNA-Binding Proteins, Transcription Factors, Tryptophan.
- chemical , metabolism : RNA.
- chemical , pharmacology : Guanidine.
- chemical : Bacterial Proteins.
- metabolism : Bacillus subtilis, Escherichia coli.
- Circular Dichroism, Dose-Response Relationship, Drug, Kinetics, Models, Chemical, Mutation, Protein Binding, Protein Conformation, Protein Denaturation, Protein Folding, Protein Structure, Secondary, Time Factors, Ultracentrifugation.
Abstract
TRAP (trp RNA-binding attenuation protein) is an RNA-binding protein that regulates expression of the tryptophan biosynthetic genes in Bacillus subtilis by binding to RNA targets that contain multiple GAG and UAG repeats. TRAP is composed of 11 identical subunits arranged symmetrically in a ring. The secondary structure of the protein consists entirely of antiparallel beta-sheets, beta-turns, and loops. We show here that the TRAP 11-mer can be reversibly denatured into unfolded monomers by guanidine hydrochloride. Removing the denaturant allows the protein to spontaneously renature into fully functional 11-mers. Based on this finding, we developed a subunit mixing method to hybridize wild-type and mutant subunits into heteromeric 11-mers by denaturation followed by subunit mixing renaturation. This method allows the study of subunit cooperativity in protein-ligand interaction such as RNA binding. Our data further support and extend the previously proposed two-step model for RNA binding to TRAP by showing that the initiation of binding requires at least one fully active subunit in the protein combined with one fully functional repeat in the RNA. The initiation complex tethers the RNA on the protein, thus allowing cooperative interaction with the remainder of the repeats.
DOI: 10.1074/jbc.M110860200
PubMed: 11805104
Affiliations:
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Scott</name></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:11805104</idno><idno type="pmid">11805104</idno><idno type="doi">10.1074/jbc.M110860200</idno><idno type="wicri:Area/PubMed/Corpus">002523</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002523</idno><idno type="wicri:Area/PubMed/Curation">002523</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002523</idno><idno type="wicri:Area/PubMed/Checkpoint">002375</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002375</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Creating hetero-11-mers composed of wild-type and mutant subunits to study RNA binding to TRAP.</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="Scott, David J" sort="Scott, David J" uniqKey="Scott D" first="David J" last="Scott">David J. Scott</name></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 (metabolism)</term><term>Bacterial Proteins</term><term>Circular Dichroism</term><term>Dose-Response Relationship, Drug</term><term>Escherichia coli (metabolism)</term><term>Guanidine (pharmacology)</term><term>Kinetics</term><term>Models, Chemical</term><term>Mutation</term><term>Protein Binding</term><term>Protein Conformation</term><term>Protein Denaturation</term><term>Protein Folding</term><term>Protein Structure, Secondary</term><term>RNA (metabolism)</term><term>RNA-Binding Proteins (chemistry)</term><term>Time Factors</term><term>Transcription Factors (chemistry)</term><term>Tryptophan (chemistry)</term><term>Ultracentrifugation</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN (métabolisme)</term><term>Bacillus subtilis (métabolisme)</term><term>Cinétique</term><term>Conformation des protéines</term><term>Dichroïsme circulaire</term><term>Dénaturation des protéines</term><term>Escherichia coli (métabolisme)</term><term>Facteurs de transcription ()</term><term>Facteurs temps</term><term>Guanidine (pharmacologie)</term><term>Liaison aux protéines</term><term>Modèles chimiques</term><term>Mutation</term><term>Pliage des protéines</term><term>Protéines bactériennes</term><term>Protéines de liaison à l'ARN ()</term><term>Relation dose-effet des médicaments</term><term>Structure secondaire des protéines</term><term>Tryptophane ()</term><term>Ultracentrifugation</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>RNA-Binding Proteins</term><term>Transcription Factors</term><term>Tryptophan</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>RNA</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Guanidine</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Bacterial Proteins</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Bacillus subtilis</term><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN</term><term>Bacillus subtilis</term><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="pharmacologie" xml:lang="fr"><term>Guanidine</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Circular Dichroism</term><term>Dose-Response Relationship, Drug</term><term>Kinetics</term><term>Models, Chemical</term><term>Mutation</term><term>Protein Binding</term><term>Protein Conformation</term><term>Protein Denaturation</term><term>Protein Folding</term><term>Protein Structure, Secondary</term><term>Time Factors</term><term>Ultracentrifugation</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Cinétique</term><term>Conformation des protéines</term><term>Dichroïsme circulaire</term><term>Dénaturation des protéines</term><term>Facteurs de transcription</term><term>Facteurs temps</term><term>Liaison aux protéines</term><term>Modèles chimiques</term><term>Mutation</term><term>Pliage des protéines</term><term>Protéines bactériennes</term><term>Protéines de liaison à l'ARN</term><term>Relation dose-effet des médicaments</term><term>Structure secondaire des protéines</term><term>Tryptophane</term><term>Ultracentrifugation</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">TRAP (trp RNA-binding attenuation protein) is an RNA-binding protein that regulates expression of the tryptophan biosynthetic genes in Bacillus subtilis by binding to RNA targets that contain multiple GAG and UAG repeats. TRAP is composed of 11 identical subunits arranged symmetrically in a ring. The secondary structure of the protein consists entirely of antiparallel beta-sheets, beta-turns, and loops. We show here that the TRAP 11-mer can be reversibly denatured into unfolded monomers by guanidine hydrochloride. Removing the denaturant allows the protein to spontaneously renature into fully functional 11-mers. Based on this finding, we developed a subunit mixing method to hybridize wild-type and mutant subunits into heteromeric 11-mers by denaturation followed by subunit mixing renaturation. This method allows the study of subunit cooperativity in protein-ligand interaction such as RNA binding. Our data further support and extend the previously proposed two-step model for RNA binding to TRAP by showing that the initiation of binding requires at least one fully active subunit in the protein combined with one fully functional repeat in the RNA. The initiation complex tethers the RNA on the protein, thus allowing cooperative interaction with the remainder of the repeats.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">11805104</PMID><DateCompleted><Year>2002</Year><Month>05</Month><Day>13</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Print">0021-9258</ISSN><JournalIssue CitedMedium="Print"><Volume>277</Volume><Issue>14</Issue><PubDate><Year>2002</Year><Month>Apr</Month><Day>05</Day></PubDate></JournalIssue><Title>The Journal of biological chemistry</Title><ISOAbbreviation>J. Biol. Chem.</ISOAbbreviation></Journal><ArticleTitle>Creating hetero-11-mers composed of wild-type and mutant subunits to study RNA binding to TRAP.</ArticleTitle><Pagination><MedlinePgn>11838-44</MedlinePgn></Pagination><Abstract><AbstractText>TRAP (trp RNA-binding attenuation protein) is an RNA-binding protein that regulates expression of the tryptophan biosynthetic genes in Bacillus subtilis by binding to RNA targets that contain multiple GAG and UAG repeats. TRAP is composed of 11 identical subunits arranged symmetrically in a ring. The secondary structure of the protein consists entirely of antiparallel beta-sheets, beta-turns, and loops. We show here that the TRAP 11-mer can be reversibly denatured into unfolded monomers by guanidine hydrochloride. Removing the denaturant allows the protein to spontaneously renature into fully functional 11-mers. Based on this finding, we developed a subunit mixing method to hybridize wild-type and mutant subunits into heteromeric 11-mers by denaturation followed by subunit mixing renaturation. This method allows the study of subunit cooperativity in protein-ligand interaction such as RNA binding. Our data further support and extend the previously proposed two-step model for RNA binding to TRAP by showing that the initiation of binding requires at least one fully active subunit in the protein combined with one fully functional repeat in the RNA. The initiation complex tethers the RNA on the protein, thus allowing cooperative interaction with the remainder of the repeats.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Li</LastName><ForeName>Pan T X</ForeName><Initials>PT</Initials><AffiliationInfo><Affiliation>Department of Biological Sciences, State University of New York, Buffalo, New York 14260, USA.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Scott</LastName><ForeName>David J</ForeName><Initials>DJ</Initials></Author><Author ValidYN="Y"><LastName>Gollnick</LastName><ForeName>Paul</ForeName><Initials>P</Initials></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>GM62750</GrantID><Acronym>GM</Acronym><Agency>NIGMS NIH HHS</Agency><Country>United States</Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013486">Research Support, U.S. Gov't, Non-P.H.S.</PublicationType><PublicationType UI="D013487">Research Support, U.S. Gov't, P.H.S.</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2002</Year><Month>01</Month><Day>22</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J Biol Chem</MedlineTA><NlmUniqueID>2985121R</NlmUniqueID><ISSNLinking>0021-9258</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D001426">Bacterial Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C079092">MtrB protein, Bacteria</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D016601">RNA-Binding Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D014157">Transcription Factors</NameOfSubstance></Chemical><Chemical><RegistryNumber>63231-63-0</RegistryNumber><NameOfSubstance UI="D012313">RNA</NameOfSubstance></Chemical><Chemical><RegistryNumber>8DUH1N11BX</RegistryNumber><NameOfSubstance UI="D014364">Tryptophan</NameOfSubstance></Chemical><Chemical><RegistryNumber>JU58VJ6Y3B</RegistryNumber><NameOfSubstance UI="D019791">Guanidine</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D001412" MajorTopicYN="N">Bacillus subtilis</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D001426" MajorTopicYN="Y">Bacterial Proteins</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002942" MajorTopicYN="N">Circular Dichroism</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004305" MajorTopicYN="N">Dose-Response Relationship, Drug</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004926" MajorTopicYN="N">Escherichia coli</DescriptorName><QualifierName UI="Q000378" 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UI="D017433" MajorTopicYN="N">Protein Structure, Secondary</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D012313" MajorTopicYN="N">RNA</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016601" MajorTopicYN="N">RNA-Binding Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013997" MajorTopicYN="N">Time Factors</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D014157" MajorTopicYN="N">Transcription Factors</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="Y">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014364" MajorTopicYN="N">Tryptophan</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D014461" MajorTopicYN="N">Ultracentrifugation</DescriptorName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2002</Year><Month>1</Month><Day>24</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2002</Year><Month>5</Month><Day>15</Day><Hour>10</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2002</Year><Month>1</Month><Day>24</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">11805104</ArticleId><ArticleId IdType="doi">10.1074/jbc.M110860200</ArticleId><ArticleId IdType="pii">M110860200</ArticleId></ArticleIdList></PubmedData></pubmed><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><name sortKey="Scott, David J" sort="Scott, David J" uniqKey="Scott D" first="David J" last="Scott">David J. Scott</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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