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Title: Proteome investigation of the global regulatory role of σ54 in response to gentisate induction in Pseudomonas alcaligenes NCIMB 9867
Authors: Zhao, B.
Yeo, C.C.
Poh, C.L. 
Keywords: Gentisate pathway
Pseudomonas alcaligenes
Sigma 54
Issue Date: May-2005
Citation: Zhao, B., Yeo, C.C., Poh, C.L. (2005-05). Proteome investigation of the global regulatory role of σ54 in response to gentisate induction in Pseudomonas alcaligenes NCIMB 9867. Proteomics 5 (7) : 1868-1876. ScholarBank@NUS Repository.
Abstract: Pseudomonas alcaligenes NCIMB 9867 (strain P25X) utilizes the gentisate pathway for the degradation of aromatic hydrocarbons. The gene encoding the alternative sigma (σ) factor σ54, rpoN, was cloned from strain P25X and a rpoN knock-out strain, designated G54, was constructed by insertional inactivation with a kanamycin resistance gene cassette. The role of σ54 in the physiological response of P. alcaligenes P25X to gentisate induction was assessed by comparing the global protein expression profiles of the wild-type P25X with the rpoN mutant strain G54. Analysis of two-dimensional polyacrylamide gel electrophoresis gels showed that 39 out of 355 prominent protein spots exhibited differential expression as a result of the insertional inactivation of rpoN. Identification of the protein spots by matrix-assisted laser desorption/ionization-time of flight/time of flight revealed a wide diversity of proteins that are affected by the σ54 mutation, the largest group being proteins that are involved in carbon metabolism. The strictly inducible gentisate 1,2-dioxygenase, one of two isofunctional copies of the key enzyme in the gentisate pathway, and enzymes of the TCA cycle, pyruvate metabolism and gluconeogenesis were part of this group. Other proteins that are part of the σ54 regulon include enzymes implicated in nitrogen metabolism, transport proteins, stress-response proteins and proteins involved in cell motility. The results of this study showed that σ54 plays a global regulatory role in the expression of a wide variety of genes in P. alcaligenes, including the wild-type response to the presence of the aromatic inducer, gentisate. © 2005 WILEY-VCH Verlag GmbH & Co. KGaA.
Source Title: Proteomics
ISSN: 16159853
DOI: 10.1002/pmic.200401081
Appears in Collections:Staff Publications

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