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PMID: 11741861 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Domain interactions on the ntr signal transduction pathway: two-hybrid analysis of mutant and truncated derivatives of histidine kinase NtrB.

Journal of bacteriology ·Vol. 184 ·No. 1 ·2002-01-00 ·Pages 200-6

Martínez-Argudo I, Salinas P, Maldonado R, Contreras A

Abstract

We have used the yeast two-hybrid system to analyze protein-protein interactions mediated by domains of regulatory proteins of the ntr signal transduction system, including interactions among NtrB derivatives and their interactions with NtrC and PII from Klebsiella pneumoniae. Interactions took place only between proteins or protein domains belonging to the ntr signal transduction system and not between proteins or domains from noncognate regulators. NtrB and its transmitter domain, but not NtrC, CheA, or the cytoplasmic C terminus of EnvZ, interacted with PII. In addition, interaction of NtrB with NtrC, but not with PII, depended on the histidine phosphotransfer domain. Point mutation A129T, diminishing the NtrC phosphatase activity of NtrB, affected the strength of the signals between NtrC and the transmitter module of NtrB but had no impact on PII signals, suggesting that A129T prevents the conformational change needed by NtrB to function as a phosphatase for NtrC, rather than disturbing binding to PII.

MeSH Terms
Bacterial Proteins Catalytic Domain DNA-Binding Proteins/metabolism Histidine Kinase Klebsiella pneumoniae/metabolism Mutation Nucleotidyltransferases/metabolism PII Nitrogen Regulatory Proteins Phosphoprotein Phosphatases/genetics,metabolism Protein Binding Protein Kinases/genetics,metabolism Protein Structure, Tertiary Sequence Deletion Signal Transduction Trans-Activators Transcription Factors Two-Hybrid System Techniques
Chemicals
Bacterial Proteins DNA-Binding Proteins PII Nitrogen Regulatory Proteins Trans-Activators Transcription Factors Protein Kinases Histidine Kinase protein kinase-phosphatase NTRB Nucleotidyltransferases regulatory protein uridylyltransferase Phosphoprotein Phosphatases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Martínez-Argudo Isabel
División de Genética, Universidad de Alicante, Apartado 99, E-03080 Alicante, Spain.
Salinas Paloma
Maldonado Rafael
Contreras Asunción
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.1.200-206.2002
Published
2002-01-00
Pages
200-6
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC134775
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.184.1.200-206.2002 PMC Full Text View journal details
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