1887

Abstract

In serovar Typhimurium, the genomic island GEI4417/4436 has recently been identified to be responsible for -inositol (MI) utilization. Here, two of the four island-encoded permeases are identified as the MI transporters of this pathogen. In-frame deletion of (STM4418) led to a severe growth defect, and deletion of (STM4419) to a slight growth defect in the presence of MI. These phenotypes could be complemented by providing the putative transporter genes Bioluminescence-based reporter assays demonstrated a strong induction of their promoters P and P in the presence of MI but not of glucose. Deletion of , which encodes the negative regulator of most genes involved in MI degradation, resulted in upregulation of P and P, indicating that the expression of IolT1 and IolT2 is repressed by IolR. This finding was supported by bandshift assays using purified IolR. Both transporters are located in the membrane when expressed in . Heterologously expressed IolT1 had its optimal activity at pH 5.5. Together with the strongly reduced MI uptake in the presence of protonophores, this indicates that IolT1 operates as a proton symporter. Using -[1,2-[H](N)]inositol, a saturable uptake activity of IolT1 with a value between 0.49 and 0.79 mM was determined in DH5 expressing IolT1, in serovar Typhimurium strain 14028, and in mutant 14028 Δ. Phylogenetic analysis of IolT1 identified putative MI transporters in Gram-negative bacteria also able to utilize MI.

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2010-01-01
2024-03-29
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