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Journal of Bacteriology, January 1999, p. 368-374, Vol. 181, No. 2
Department of Bacteriology, University of
Wisconsin
Received 3 August 1998/Accepted 3 November 1998
The eutF locus of Salmonella typhimurium
LT2 was identified as a locus necessary for the utilization of
ethanolamine as a sole carbon source. Initial models suggested that
EutF was involved in either ethanolamine transport or was a
transcriptional regulator of an ethanolamine transporter. Phenotypic
characterization of eutF mutants suggested EutF was somehow
involved in 1,2-propanediol, propionate, and succinate utilization.
Here we provide evidence that two alleles defining the eutF
locus,
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Molecular Characterization of eutF Mutants of
Salmonella typhimurium LT2 Identifies eutF
Lesions as Partial-Loss-of-Function tonB Alleles
and
Madison, Madison, Wisconsin 53706-1567
903 and eutF1115, are partial-loss-of-function tonB alleles. Both mutations were
complemented by plasmids containing a wild-type allele of the
Escherichia coli tonB gene. Immunoblot analysis using TonB
monoclonal antibodies detected a TonB fusion protein in strains
carrying eutF alleles. Molecular analysis of the
903 allele identified a deletion that resulted in the
fusion of the 3' end of tonB with the 3' end of trpA. In-frame translation of the tonB-trpA
fusion resulted in the final 9 amino acids of TonB being replaced by a
45-amino-acid addition. We isolated a derivative of a strain carrying
allele
903 that regained the ability to grow on
ethanolamine as a carbon and energy source. The molecular
characterization of the mutation that corrected the Eut
phenotype caused by allele
903 showed that the new
mutation was a deletion of two nucleotides at the tonB-trpA
fusion site. This deletion resulted in a frameshift that replaced the
45-amino-acid addition with a 5-amino-acid addition. This change
resulted in a TonB protein with sufficient activity to restore growth
on ethanolamine and eut operon expression to nearly
wild-type levels. It was concluded that the observed EutF phenotypes
were due to the partial loss of TonB function, which is proposed to
result in reduced cobalamin and ferric siderophore transport in an
aerobic environment; thus, the eutF locus does not exist.
*
Corresponding author. Mailing address: Department of
Bacteriology, University of Wisconsin
Madison, 1550 Linden Dr.,
Madison, WI 53706-1567. Phone: (608) 262-7379. Fax: (608) 262-9865. E-mail: jcescala{at}facstaff.wisc.edu.
Present address: Department of Microbiology and Molecular Genetics,
Harvard Medical School, Boston, Mass.
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