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PMID: 10671442 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Characterization and role of tbuX in utilization of toluene by Ralstonia pickettii PKO1.

Journal of bacteriology ·Vol. 182 ·No. 5 ·2000-03-00 ·Pages 1232-42

Kahng HY, Byrne AM, Olsen RH, Kukor JJ

Abstract

The tbu regulon of Ralstonia pickettii PKO1 encodes enzymes involved in the catabolism of toluene, benzene, and related alkylaromatic hydrocarbons. The first operon in this regulon contains genes that encode the tbu pathway's initial catabolic enzyme, toluene-3-monooxygenase, as well as TbuT, the NtrC-like transcriptional activator for the entire regulon. It has been previously shown that the organization of tbuT, which is located immediately downstream of tbuA1UBVA2C, and the associated promoter (PtbuA1) is unique in that it results in a cascade type of up-regulation of tbuT in response to a variety of effector compounds. In our efforts to further characterize this unusual mode of gene regulation, we discovered another open reading frame, encoded on the strand opposite that of tbuT, 63 bp downstream of the tbuT stop codon. The 1,374-bp open reading frame, encoding a 458-amino-acid peptide, was designated tbuX. The predicted amino acid sequence of TbuX exhibited significant similarity to several putative outer membrane proteins from aromatic hydrocarbon-degrading bacteria, as well as to FadL, an outer membrane protein needed for uptake of long-chain fatty acids in Escherichia coli. Based on sequence analysis, transcriptional and expression studies, and deletion analysis, TbuX seems to play an important role in the catabolism of toluene in R. pickettii PKO1. In addition, the expression of tbuX appears to be regulated in a manner such that low levels of TbuX are always present within the cell, whereas upon toluene exposure these levels dramatically increase, even more than those of toluene-3-monooxygenase. This expression pattern may relate to the possible role of TbuX as a facilitator of toluene entry into the cell.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,physiology Base Sequence DNA, Bacterial/chemistry,genetics Gene Deletion Gene Expression Regulation, Bacterial Genes, Bacterial/genetics Gram-Negative Aerobic Rods and Cocci/chemistry,genetics Molecular Sequence Data Promoter Regions, Genetic/genetics Regulatory Sequences, Nucleic Acid Regulon/genetics Sequence Analysis, DNA Sequence Homology, Amino Acid Sequence Homology, Nucleic Acid Toluene/metabolism Transcription, Genetic
Chemicals
Bacterial Proteins DNA, Bacterial TbuX protein, Burkholderia pickettii Toluene
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kahng H Y
Biotechnology Center for Agriculture and the Environment, Rutgers University, New Brunswick, New Jersey 08901-8520, USA.
Byrne A M
Olsen R H
Kukor J J
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.182.5.1232-1242.2000
Published
2000-03-00
Pages
1232-42
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC94407
Subset
IM
Grants
NCRR NIH HHS · M01 RR000042 · United States
NIEHS NIH HHS · P42 ES004911 · United States
NCRR NIH HHS · M01RR00042 · United States
NIEHS NIH HHS · P42-ES-04911 · United States
Databases
GENBANK
AF100782
External Links
PubMed source DOI: 10.1128/JB.182.5.1232-1242.2000 PMC Full Text View journal details
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