Document Detail


A functional gltB gene is essential for utilization of acidic amino acids and expression of periplasmic glutaminase/asparaginase (PGA) by Pseudomonas putida KT2440.
MedLine Citation:
PMID:  14624355     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Pseudomonas putida KT2440, a root-colonizing fluorescent pseudomonad, is capable of utilizing acidic amino acids (Asp and Glu) and their amides (Asn and Gln) as its sole source of carbon and nitrogen. The uptake of Gln and Asn is facilitated by a periplasmic glutaminase/asparaginase (PGA), which hydrolyses Asn and Gln to the respective dicarboxylates. Here, we describe transposon mutagenesis of P. putida KT2440 with a self-cloning promoter probe vector, Tn 5-OT182. Transconjugants defective in Glu-mediated PGA induction were selected for further studies. In most clones the transposon was found to have integrated into the gltB gene, which encodes the major subunit of the glutamate synthase (GOGAT). The transconjugants were nonmotile, no longer showed a chemotactic response towards amino acids, and could not survive prolonged periods of starvation. The acidic amino acids and their amides supported growth of the transconjugants only when supplied together with glucose, suggesting that the gltB-mutants had lost the ability to utilize amino acids as a carbon source. To confirm that gltB inactivation was the cause of this phenotype, we constructed a mutant with a targeted disruption of gltB. This strain behaved like the clones obtained by random mutagenesis, and failed to express not only PGA but also a number of other Glu-induced proteins. In contrast to wild-type cells, the gltB(-) strain accumulated considerable amounts of both Glu and Gln during long-term incubation.
Authors:
A M Sonawane; K H Röhm
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2003-11-18
Journal Detail:
Title:  Molecular genetics and genomics : MGG     Volume:  271     ISSN:  1617-4615     ISO Abbreviation:  Mol. Genet. Genomics     Publication Date:  2004 Feb 
Date Detail:
Created Date:  2004-02-17     Completed Date:  2004-04-13     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  101093320     Medline TA:  Mol Genet Genomics     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  33-9     Citation Subset:  IM    
Affiliation:
Institute of Physiological Chemistry, Philipps University, 35033 Marburg, Germany. avinash@mailer.uni-marburg.de
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MeSH Terms
Descriptor/Qualifier:
Amidohydrolases / genetics*,  metabolism*
Amino Acids, Acidic / metabolism*
Base Sequence
Cell Division
Chemotaxis
DNA, Bacterial / genetics
Gene Expression
Genes, Bacterial*
Movement
Mutagenesis, Insertional
Phenotype
Pseudomonas putida / genetics*,  metabolism*
Chemical
Reg. No./Substance:
0/Amino Acids, Acidic; 0/DNA, Bacterial; EC 3.5.-/Amidohydrolases; EC 3.5.1.38/glutamin-(asparagin-)ase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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