PMID:7009581

Citation	Lazzaroni, JC and Portalier, RC (1981) Genetic and biochemical characterization of periplasmic-leaky mutants of Escherichia coli K-12. J. Bacteriol. 145:1351-8
Abstract	Periplasmic-leaky mutants of Escherichia coli K-12 were isolated after nitrosoguanidine-induced mutagenesis. They released periplasmic enzymes into the extracellular medium. Excretion of alkaline phosphatase, which started immediately in the early exponential phase of growth, could reach up to 90% of the total enzyme production in the stationary phase. Leaky mutants were sensitive to ethylenediaminetetraacetic acid, cholic acid, and the antibiotics rifampin, chloramphenicol, mitomycin C, and ampicillin. Furthermore, they were resistant to colicin E1 and partially resistant to phage TuLa. Their genetic characterization showed that the lky mutations mapped between the suc and gal markers, near or in the tolPAB locus. A biochemical analysis of cell envelope components showed that periplasmic-leaky mutants contained reduced amounts of major outer membrane protein OmpF and increased amounts of a 16,000-dalton outer membrane protein.
Links	PubMed PMC217139
Keywords	Alkaline Phosphatase/metabolism; Bacterial Proteins/metabolism; Cell Membrane/physiology; Chromosome Mapping; Chromosomes, Bacterial; Escherichia coli/genetics; Escherichia coli/physiology; Genes; Membrane Proteins/metabolism; Mutation
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Materials and Methods Used

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Phenotype Annotations

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Phenotype of	Taxon Information	Genotype Information (if known)	Condition Information	OMP ID	OMP Term Name	ECO ID	ECO Term Name	Notes	Status
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: Gal5 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207	Reference Condition:	0000085	cell envelope phenotype	0000178	in vivo assay evidence	A mutation in the strain 207's tolPAB operon caused the exretion of alakaline phosphate, RNAase I and DNAse I.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: Gal5 NCBI Taxon ID: 83333	Genotype of Reference Strain: phosT+ Genotype of Experimental Strain : phosT- in 188	Reference Condition:			0000178	in vivo assay evidence	Mutation the phosT gene caused arsenate resistance.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: Gal5 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 236	Reference Condition:	0000085	cell envelope phenotype	0000178	in vivo assay evidence	A mutation in the strain 236's tolPAB operon caused the exretion of alakaline phosphate, RNAase I and DNAse I.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 207 NCBI Taxon ID: 83333	Genotype of Reference Strain: phosT+ Genotype of Experimental Strain : phosT- in 207c	Reference Condition:			0000178	in vivo assay evidence	Mutation the phosT gene caused arsenate resistance.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 236c	Reference Condition:			0000178	in vivo assay evidence	Mutation the tolPAB gene caused increased activity of alkaline phosphatase. See Figure 1.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 236 NCBI Taxon ID: 83333	Genotype of Reference Strain: phosT+ Genotype of Experimental Strain : phosT- in 236c	Reference Condition:			0000178	in vivo assay evidence	Mutation the phosT gene caused arsenate resistance.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	Mutation the tolPAB gene caused increased activity of alkaline phosphatase. See Figure 1.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The distribution of alkaline phosphatase activity within perplasmic leaky mutants showed that the remaining intracellular enzyme could be released by osmotic shock.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAP- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon conferred resistance toward the phage TuIa.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 236c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon conferred resistance toward the phage TuIa.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased sensitivity to rifampin See table 5. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased resistance toward SDS. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: Gal5 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207	Reference Condition:					Analysis by electrophoresis with SDS-polyacrylamide slag gels reviled that phosphate-regulated periplasmic protiens P1( alkaline phosphatase, monomer), P2 & P4(a phosphate binding protein) were released by depressed Mutant 207 cells. See figure 2 for experimental results.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	Mutation to the tolPAB caused mutation caused the excretion of protiens P1 and P2 and the absence of band P4 (thought to be due to a phoS mutation).
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased sensitivity to mitomycin C See table 5. The same was seen ins strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased sensitivity to deoxycholic acid See table 5. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased sensitivity to EDTA See table 5. The same was seen ins strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased resistance Triton X-100. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased resistance toward acridine organge. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased resistance toward Brij 58. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:			0000178	in vivo assay evidence	The mutation to the tolPAB operon caused increased resistance toward methylen blue. The same was seen in strain 236c.
a mutation or genetic difference within a strain	Taxon: Escherichia coli Strain: K-12 Substrain: 188 NCBI Taxon ID: 83333	Genotype of Reference Strain: tolPAB+ Genotype of Experimental Strain : tolPAB- in 207c	Reference Condition:					The mutation to the tolPAB operon caused increased resistance toward SDS, Triton X-100, Brij 58, acridine organge, methylen blue and crystal violet. The same was seen in strain 236c.
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Notes

• It should be noted that the operon tolPAB is listed in the phenotype table due to the fact the exact gene mutated is unclear.
• phoS is a synonym of pstS

References

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