Amino Acids Important for DNA Recognition by the Response Regulator OmpRS

Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A det...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:The Journal of biological chemistry 2008-03, Vol.283 (13), p.8664-8677
Hauptverfasser: Rhee, Jee Eun, Sheng, Wanyun, Morgan, Leslie K., Nolet, Ryan, Liao, Xiubei, Kenney, Linda J.
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
container_end_page 8677
container_issue 13
container_start_page 8664
container_title The Journal of biological chemistry
container_volume 283
creator Rhee, Jee Eun
Sheng, Wanyun
Morgan, Leslie K.
Nolet, Ryan
Liao, Xiubei
Kenney, Linda J.
description Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A detailed mechanistic picture of how OmpR binds to DNA and activates transcription is lacking. We used NMR spectroscopy to solve the solution structure of the C-terminal domain of OmpR (OmpR C ) and to analyze the chemical shift changes that occur upon DNA binding. There is little overlap in the interaction surface with residues of PhoB that were reportedly involved in protein/protein interactions in its head-to-tail dimer. Multiple factors account for the lack of overlap. One is that the spacing between the OmpR half-sites is shorter than observed with PhoB, requiring the arrangement of the two OmpR molecules to be different from that of the PhoB dimer on DNA. A second is the demonstration herein that OmpR can bind to its high affinity site as a monomer. As a result, OmpR C appears to be capable of adopting alternative orientations depending on the precise base composition of the binding site, which also contributes to the lack of overlap. In the presence of DNA, chemical shift changes occur in OmpR in the recognition α-helix 3, the loop between β-strand 4 and α-helix 1, and the loop between β-strands 5 and 6. DNA contact residues are Val 203 (T), Arg 207 (G), and Arg 209 (phosphate backbone). Our results suggest that OmpR binds to DNA as a monomer and then forms a symmetric or asymmetric dimer, depending on the binding site. We propose that during activation OmpR binds to DNA and undergoes a conformational change that promotes phosphorylation of the N-terminal receiver domain, the receiver domains dimerize, and then the second monomer binds to DNA. The flexible linker of OmpR enables the second monomer to bind in multiple orientations (head-to-tail and head-to-head), depending on the specific DNA contacts.
doi_str_mv 10.1074/jbc.M705550200
format Article
fullrecord <record><control><sourceid>pubmedcentral</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2417188</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>pubmedcentral_primary_oai_pubmedcentral_nih_gov_2417188</sourcerecordid><originalsourceid>FETCH-pubmedcentral_primary_oai_pubmedcentral_nih_gov_24171883</originalsourceid><addsrcrecordid>eNqljD1PwzAYhC1URAPtyuw_kPK-Saw4C1LEh-gASIWBzXJSN3UVf8h2kfrvCRILc2-503OnI-QWYYVQV3eHrl-91sAYgwLggmQIvMxLhl8zkgEUmDcF43NyHeMBJlUNXpE5cmwYIM_IujXaOtr2ehvp2ngXkrSJ7lygj28t3ajeDVYn7SztTjTt1YSidzb-huE4yjQt343ffCzI5U6OUS3__IbcPz99Przk_tgZte2VTUGOwgdtZDgJJ7X431i9F4P7FkWFNXJenn3wA7qDWto</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype></control><display><type>article</type><title>Amino Acids Important for DNA Recognition by the Response Regulator OmpRS</title><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><creator>Rhee, Jee Eun ; Sheng, Wanyun ; Morgan, Leslie K. ; Nolet, Ryan ; Liao, Xiubei ; Kenney, Linda J.</creator><creatorcontrib>Rhee, Jee Eun ; Sheng, Wanyun ; Morgan, Leslie K. ; Nolet, Ryan ; Liao, Xiubei ; Kenney, Linda J.</creatorcontrib><description>Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A detailed mechanistic picture of how OmpR binds to DNA and activates transcription is lacking. We used NMR spectroscopy to solve the solution structure of the C-terminal domain of OmpR (OmpR C ) and to analyze the chemical shift changes that occur upon DNA binding. There is little overlap in the interaction surface with residues of PhoB that were reportedly involved in protein/protein interactions in its head-to-tail dimer. Multiple factors account for the lack of overlap. One is that the spacing between the OmpR half-sites is shorter than observed with PhoB, requiring the arrangement of the two OmpR molecules to be different from that of the PhoB dimer on DNA. A second is the demonstration herein that OmpR can bind to its high affinity site as a monomer. As a result, OmpR C appears to be capable of adopting alternative orientations depending on the precise base composition of the binding site, which also contributes to the lack of overlap. In the presence of DNA, chemical shift changes occur in OmpR in the recognition α-helix 3, the loop between β-strand 4 and α-helix 1, and the loop between β-strands 5 and 6. DNA contact residues are Val 203 (T), Arg 207 (G), and Arg 209 (phosphate backbone). Our results suggest that OmpR binds to DNA as a monomer and then forms a symmetric or asymmetric dimer, depending on the binding site. We propose that during activation OmpR binds to DNA and undergoes a conformational change that promotes phosphorylation of the N-terminal receiver domain, the receiver domains dimerize, and then the second monomer binds to DNA. The flexible linker of OmpR enables the second monomer to bind in multiple orientations (head-to-tail and head-to-head), depending on the specific DNA contacts.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M705550200</identifier><identifier>PMID: 18195018</identifier><language>eng</language><publisher>American Society for Biochemistry and Molecular Biology</publisher><subject>Transcription, Chromatin, and Epigenetics</subject><ispartof>The Journal of biological chemistry, 2008-03, Vol.283 (13), p.8664-8677</ispartof><rights>Copyright © 2008, The American Society for Biochemistry and Molecular Biology, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2417188/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2417188/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,53791,53793</link.rule.ids></links><search><creatorcontrib>Rhee, Jee Eun</creatorcontrib><creatorcontrib>Sheng, Wanyun</creatorcontrib><creatorcontrib>Morgan, Leslie K.</creatorcontrib><creatorcontrib>Nolet, Ryan</creatorcontrib><creatorcontrib>Liao, Xiubei</creatorcontrib><creatorcontrib>Kenney, Linda J.</creatorcontrib><title>Amino Acids Important for DNA Recognition by the Response Regulator OmpRS</title><title>The Journal of biological chemistry</title><description>Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A detailed mechanistic picture of how OmpR binds to DNA and activates transcription is lacking. We used NMR spectroscopy to solve the solution structure of the C-terminal domain of OmpR (OmpR C ) and to analyze the chemical shift changes that occur upon DNA binding. There is little overlap in the interaction surface with residues of PhoB that were reportedly involved in protein/protein interactions in its head-to-tail dimer. Multiple factors account for the lack of overlap. One is that the spacing between the OmpR half-sites is shorter than observed with PhoB, requiring the arrangement of the two OmpR molecules to be different from that of the PhoB dimer on DNA. A second is the demonstration herein that OmpR can bind to its high affinity site as a monomer. As a result, OmpR C appears to be capable of adopting alternative orientations depending on the precise base composition of the binding site, which also contributes to the lack of overlap. In the presence of DNA, chemical shift changes occur in OmpR in the recognition α-helix 3, the loop between β-strand 4 and α-helix 1, and the loop between β-strands 5 and 6. DNA contact residues are Val 203 (T), Arg 207 (G), and Arg 209 (phosphate backbone). Our results suggest that OmpR binds to DNA as a monomer and then forms a symmetric or asymmetric dimer, depending on the binding site. We propose that during activation OmpR binds to DNA and undergoes a conformational change that promotes phosphorylation of the N-terminal receiver domain, the receiver domains dimerize, and then the second monomer binds to DNA. The flexible linker of OmpR enables the second monomer to bind in multiple orientations (head-to-tail and head-to-head), depending on the specific DNA contacts.</description><subject>Transcription, Chromatin, and Epigenetics</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><recordid>eNqljD1PwzAYhC1URAPtyuw_kPK-Saw4C1LEh-gASIWBzXJSN3UVf8h2kfrvCRILc2-503OnI-QWYYVQV3eHrl-91sAYgwLggmQIvMxLhl8zkgEUmDcF43NyHeMBJlUNXpE5cmwYIM_IujXaOtr2ehvp2ngXkrSJ7lygj28t3ajeDVYn7SztTjTt1YSidzb-huE4yjQt343ffCzI5U6OUS3__IbcPz99Przk_tgZte2VTUGOwgdtZDgJJ7X431i9F4P7FkWFNXJenn3wA7qDWto</recordid><startdate>20080328</startdate><enddate>20080328</enddate><creator>Rhee, Jee Eun</creator><creator>Sheng, Wanyun</creator><creator>Morgan, Leslie K.</creator><creator>Nolet, Ryan</creator><creator>Liao, Xiubei</creator><creator>Kenney, Linda J.</creator><general>American Society for Biochemistry and Molecular Biology</general><scope>5PM</scope></search><sort><creationdate>20080328</creationdate><title>Amino Acids Important for DNA Recognition by the Response Regulator OmpRS</title><author>Rhee, Jee Eun ; Sheng, Wanyun ; Morgan, Leslie K. ; Nolet, Ryan ; Liao, Xiubei ; Kenney, Linda J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-pubmedcentral_primary_oai_pubmedcentral_nih_gov_24171883</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Transcription, Chromatin, and Epigenetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rhee, Jee Eun</creatorcontrib><creatorcontrib>Sheng, Wanyun</creatorcontrib><creatorcontrib>Morgan, Leslie K.</creatorcontrib><creatorcontrib>Nolet, Ryan</creatorcontrib><creatorcontrib>Liao, Xiubei</creatorcontrib><creatorcontrib>Kenney, Linda J.</creatorcontrib><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rhee, Jee Eun</au><au>Sheng, Wanyun</au><au>Morgan, Leslie K.</au><au>Nolet, Ryan</au><au>Liao, Xiubei</au><au>Kenney, Linda J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Amino Acids Important for DNA Recognition by the Response Regulator OmpRS</atitle><jtitle>The Journal of biological chemistry</jtitle><date>2008-03-28</date><risdate>2008</risdate><volume>283</volume><issue>13</issue><spage>8664</spage><epage>8677</epage><pages>8664-8677</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Response regulators undergo regulated phosphorylation and dephosphorylation at conserved aspartic acid residues in bacterial signal transduction systems. OmpR is a winged helix-turnhelix DNA-binding protein that functions as a global regulator in bacteria and is also important in pathogenesis. A detailed mechanistic picture of how OmpR binds to DNA and activates transcription is lacking. We used NMR spectroscopy to solve the solution structure of the C-terminal domain of OmpR (OmpR C ) and to analyze the chemical shift changes that occur upon DNA binding. There is little overlap in the interaction surface with residues of PhoB that were reportedly involved in protein/protein interactions in its head-to-tail dimer. Multiple factors account for the lack of overlap. One is that the spacing between the OmpR half-sites is shorter than observed with PhoB, requiring the arrangement of the two OmpR molecules to be different from that of the PhoB dimer on DNA. A second is the demonstration herein that OmpR can bind to its high affinity site as a monomer. As a result, OmpR C appears to be capable of adopting alternative orientations depending on the precise base composition of the binding site, which also contributes to the lack of overlap. In the presence of DNA, chemical shift changes occur in OmpR in the recognition α-helix 3, the loop between β-strand 4 and α-helix 1, and the loop between β-strands 5 and 6. DNA contact residues are Val 203 (T), Arg 207 (G), and Arg 209 (phosphate backbone). Our results suggest that OmpR binds to DNA as a monomer and then forms a symmetric or asymmetric dimer, depending on the binding site. We propose that during activation OmpR binds to DNA and undergoes a conformational change that promotes phosphorylation of the N-terminal receiver domain, the receiver domains dimerize, and then the second monomer binds to DNA. The flexible linker of OmpR enables the second monomer to bind in multiple orientations (head-to-tail and head-to-head), depending on the specific DNA contacts.</abstract><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>18195018</pmid><doi>10.1074/jbc.M705550200</doi></addata></record>
fulltext fulltext
identifier ISSN: 0021-9258
ispartof The Journal of biological chemistry, 2008-03, Vol.283 (13), p.8664-8677
issn 0021-9258
1083-351X
language eng
recordid cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2417188
source EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection
subjects Transcription, Chromatin, and Epigenetics
title Amino Acids Important for DNA Recognition by the Response Regulator OmpRS
url https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T01%3A49%3A20IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-pubmedcentral&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Amino%20Acids%20Important%20for%20DNA%20Recognition%20by%20the%20Response%20Regulator%20OmpRS&rft.jtitle=The%20Journal%20of%20biological%20chemistry&rft.au=Rhee,%20Jee%20Eun&rft.date=2008-03-28&rft.volume=283&rft.issue=13&rft.spage=8664&rft.epage=8677&rft.pages=8664-8677&rft.issn=0021-9258&rft.eissn=1083-351X&rft_id=info:doi/10.1074/jbc.M705550200&rft_dat=%3Cpubmedcentral%3Epubmedcentral_primary_oai_pubmedcentral_nih_gov_2417188%3C/pubmedcentral%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/18195018&rfr_iscdi=true