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J Biol Chem, Vol. 274, Issue 22, 15562-15568, May 28, 1999
From the Centro Nacional de Biotecnología del Consejo
Superior de Investigaciones Científicas,
Madrid 28049, Spain
The gene cluster adjacent to the sequence
of rpoN (encoding sigma factor
The IIANtr (PtsN) Protein of Pseudomonas
putida Mediates the C Source Inhibition of the
54-dependent Pu Promoter of
the TOL Plasmid
54) of
Pseudomonas putida has been studied with respect to the C source regulation of the Pu promoter of the
upper TOL (toluene catabolism) operon. The region includes
four open reading frames (ORFs), two of which (named ptsN
and ptsO genes) encode proteins similar to components of
the phosphoenolpyruvate:sugar phosphotransferase system. Each of the
four genes was disrupted with a nonpolar insertion, and the effects in
the inhibition caused by glucose on Pu activity were
inspected with a lacZ reporter system. Although cells
lacking ORF102, ORF284, and ptsO did not display any
evident phenotype under the conditions tested, the loss of
ptsN, which encodes the IIANtr protein, made
Pu unresponsive to repression by glucose. The
ptsN mutant had rates of glucose/gluconate consumption
identical to those of the wild type, thus ruling out indirect effects
mediated by the transport of the carbohydrate. A site-directed
ptsN mutant in which the conserved phospho-acceptor site
His68 of IIANtr was replaced by an aspartic
acid residue made Pu blind to the presence or absence of
glucose, thus supporting the notion that phosphorylation of
IIANtr mediates the C source inhibition of the promoter.
These data substantiate the existence of a molecular pathway for
co-regulation of some 54 promoters in which
IIANtr is a key protein intermediate.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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