Domain swapping reveals that the N-terminal domain of the sensor kinase KdpD in Escherichia coli is important for signaling

Publikationstyp:
Zeitschriftenaufsatz
Metadaten:
Autoren
  • Ralf Heermann
  • Marie-Luise Lippert
  • Kirsten Jung
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000268668700001&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1186/1471-2180-9-133
Externe Identifier
  • Clarivate Analytics Document Solution ID: 479OB
  • PubMed Identifier: 19589130
ISSN
1471-2180
Zeitschrift
BMC MICROBIOLOGY
Artikelnummer
ARTN 133
Datum der Veröffentlichung
2009
Status
Published
Titel
Domain swapping reveals that the N-terminal domain of the sensor kinase KdpD in <i>Escherichia coli</i> is important for signaling
Sub types
  • Article
Ausgabe der Zeitschrift
9

Datenquelle: Web of Science (Lite)

Andere Metadatenquellen:
Autoren
  • Ralf Heermann
  • Marie-Luise Lippert
  • Kirsten Jung
DOI
10.1186/1471-2180-9-133
ISSN
1471-2180
Ausgabe der Veröffentlichung
1
Zeitschrift
BMC Microbiology
Sprache
en
Paginierung
133 - 133
Datum der Veröffentlichung
2009
Status
Published
Herausgeber
Springer Science and Business Media LLC
Herausgeber URL
http://dx.doi.org/10.1186/1471-2180-9-133
Datum der Datenerfassung
2016
Titel
Domain swapping reveals that the N-terminal domain of the sensor kinase KdpD in Escherichia coli is important for signaling
Ausgabe der Zeitschrift
9

Datenquelle: Crossref

Abstract
<h4>Background</h4>The KdpD/KdpE two-component system of Escherichia coli regulates expression of the kdpFABC operon encoding the high affinity K+ transport system KdpFABC. The input domain of KdpD comprises a domain that belongs to the family of universal stress proteins (Usp). It has been previously demonstrated that UspC binds to this domain, resulting in KdpD/KdpE scaffolding under salt stress. However the mechanistic significance of this domain for signaling remains unclear. Here, we employed a "domain swapping" approach to replace the KdpD-Usp domain with four homologous domains or with the six soluble Usp proteins of E. coli.<h4>Results</h4>Full response to salt stress was only achieved with a chimera that contains UspC, probably due to unaffected scaffolding of the KdpD/KdpE signaling cascade by soluble UspC. Unexpectedly, chimeras containing either UspF or UspG not only prevented kdpFABC expression under salt stress but also under K+ limiting conditions, although these hybrid proteins exhibited kinase and phosphotransferase activities in vitro. These are the first KdpD derivatives that do not respond to K+ limitation due to alterations in the N-terminal domain. Analysis of the KdpD-Usp tertiary structure revealed that this domain has a net positively charged surface, while UspF and UspG are characterized by net negative surface charges.<h4>Conclusion</h4>The Usp domain within KdpD not only functions as a binding surface for the scaffold UspC, but it is also important for KdpD signaling. We propose that KdpD sensing/signaling involves alterations of electrostatic interactions between the large N- and C-terminal cytoplasmic domains.
Addresses
  • Ludwig-Maximilians-Universität München, Biozentrum, Bereich Mikrobiologie, D-82152 Martinsried, Germany. heermann@lmu.de
Autoren
  • Ralf Heermann
  • Marie-Luise Lippert
  • Kirsten Jung
DOI
10.1186/1471-2180-9-133
eISSN
1471-2180
Externe Identifier
  • PubMed Identifier: 19589130
  • PubMed Central ID: PMC2714519
Open access
true
ISSN
1471-2180
Zeitschrift
BMC microbiology
Schlüsselwörter
  • Escherichia coli
  • Sodium Chloride
  • Potassium
  • Protein Kinases
  • Escherichia coli Proteins
  • Heat-Shock Proteins
  • Recombinant Proteins
  • Sequence Alignment
  • Signal Transduction
  • Binding Sites
  • Amino Acid Sequence
  • Protein Structure, Tertiary
  • Phosphorylation
  • Molecular Sequence Data
  • Protein Interaction Domains and Motifs
  • Stress, Physiological
Sprache
eng
Medium
Electronic
Online publication date
2009
Open access status
Open Access
Paginierung
133
Datum der Veröffentlichung
2009
Status
Published
Publisher licence
CC BY
Datum der Datenerfassung
2009
Titel
Domain swapping reveals that the N-terminal domain of the sensor kinase KdpD in Escherichia coli is important for signaling.
Sub types
  • Research Support, Non-U.S. Gov't
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
9

Files

https://bmcmicrobiol.biomedcentral.com/counter/pdf/10.1186/1471-2180-9-133 http://www.biomedcentral.com/content/pdf/1471-2180-9-133.pdf https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19589130/pdf/?tool=EBI https://europepmc.org/articles/PMC2714519?pdf=render

Datenquelle: Europe PubMed Central

Abstract
BACKGROUND: The KdpD/KdpE two-component system of Escherichia coli regulates expression of the kdpFABC operon encoding the high affinity K+ transport system KdpFABC. The input domain of KdpD comprises a domain that belongs to the family of universal stress proteins (Usp). It has been previously demonstrated that UspC binds to this domain, resulting in KdpD/KdpE scaffolding under salt stress. However the mechanistic significance of this domain for signaling remains unclear. Here, we employed a "domain swapping" approach to replace the KdpD-Usp domain with four homologous domains or with the six soluble Usp proteins of E. coli. RESULTS: Full response to salt stress was only achieved with a chimera that contains UspC, probably due to unaffected scaffolding of the KdpD/KdpE signaling cascade by soluble UspC. Unexpectedly, chimeras containing either UspF or UspG not only prevented kdpFABC expression under salt stress but also under K+ limiting conditions, although these hybrid proteins exhibited kinase and phosphotransferase activities in vitro. These are the first KdpD derivatives that do not respond to K+ limitation due to alterations in the N-terminal domain. Analysis of the KdpD-Usp tertiary structure revealed that this domain has a net positively charged surface, while UspF and UspG are characterized by net negative surface charges. CONCLUSION: The Usp domain within KdpD not only functions as a binding surface for the scaffold UspC, but it is also important for KdpD signaling. We propose that KdpD sensing/signaling involves alterations of electrostatic interactions between the large N- and C-terminal cytoplasmic domains.
Date of acceptance
2009
Autoren
  • Ralf Heermann
  • Marie-Luise Lippert
  • Kirsten Jung
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/19589130
DOI
10.1186/1471-2180-9-133
eISSN
1471-2180
Externe Identifier
  • PubMed Central ID: PMC2714519
Zeitschrift
BMC Microbiol
Schlüsselwörter
  • Amino Acid Sequence
  • Binding Sites
  • Escherichia coli
  • Escherichia coli Proteins
  • Heat-Shock Proteins
  • Molecular Sequence Data
  • Phosphorylation
  • Potassium
  • Protein Interaction Domains and Motifs
  • Protein Kinases
  • Protein Structure, Tertiary
  • Recombinant Proteins
  • Sequence Alignment
  • Signal Transduction
  • Sodium Chloride
  • Stress, Physiological
Sprache
eng
Country
England
Paginierung
133
PII
1471-2180-9-133
Datum der Veröffentlichung
2009
Status
Published online
Datum, an dem der Datensatz öffentlich gemacht wurde
2009
Titel
Domain swapping reveals that the N-terminal domain of the sensor kinase KdpD in Escherichia coli is important for signaling.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
9

Datenquelle: PubMed

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