Abstract
Sixty-four mutants were isolated from the LT-2 wild-type strain of Salmonella typhimurium by selecting for chromate resistance. The majority of lesions were shown to lie in the cysA gene. (i) The mutants cannot take up sulfate, a finding which verifies the role of cysA in sulfate transport. In addition, 52 sulfate-transport mutants isolated without chromate selection were defective in the cysA gene. (ii) Most had less than 25% of the binding activity of the wild-type strain. (iii) Most had normal sulfite reductase (H(2)S-nicotinamide adenine dinucleotide phosphate oxidoreductase, EC 1.8.1.2) activity. (iv) Their sulfate-binding protein (binder) appears electrophoretically and immunologically normal. (v) Amber cysA mutants also make apparently normal binder in small amounts. (vi) All classical cysA mutants tested, including two with long deletions, had normal binding activity. From these observations, it is suggested that the cysA gene does not code for the binder. But many mutations in this gene reduce the binding activity in some unknown way. Other mutants, identified as cysB mutants, had neither binding nor uptake activities and their sulfite reductase activities were similarly reduced, thus confirming the regulatory role of the cysB gene. When binder was detectable, it had wild-type properties. No mutations in the binder gene were found among more than 100 mutants examined. Thus, sulfate binding has not been established as a part of sulfate transport. However, the production of binder is intimately connected with cysA, the established sulfate transport gene, and is regulated by the same mechanism that regulates both transport and the rest of the cysteine biosynthetic pathway.
MeSH Terms
Acrylates
Cell-Free System
Chromates/pharmacology
Chromosome Mapping
Culture Media
Cysteine/metabolism
Drug Resistance, Microbial
Electrophoresis
Gels
Genes
Genetic Complementation Test
Genetics, Microbial
Immunodiffusion
Mutation
Osmosis
Oxidoreductases/metabolism
Protein Binding
Salmonella typhimurium/drug effects,enzymology,growth & development,immunology,isolation & purification,metabolism
Sulfates/metabolism
Temperature
Transduction, Genetic
Chemicals
Acrylates
Chromates
Culture Media
Gels
Sulfates
Oxidoreductases
Cysteine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ota N
Galsworthy P R
Pardee A B
References (21)
21 references, click to expand
-
Purification and properties of a sulfate-binding protein from Salmonella typhimurium.
J Biol Chem. 1966 Dec 25;241(24):5886-92
PMID: 5333560
-
Positive control by the cys-3 locus in regulation of sulfur metabolism in Neurospora.
J Mol Biol. 1968 Apr 28;33(2):423-37
PMID: 5700703
-
Isolation and characterization of ribonuclease I mutants of Escherichia coli.
J Mol Biol. 1966 Mar;16(1):67-84
PMID: 5331244
-
Suppressor Mutations in Some Cystine-Requiring Mutants of Salmonella Typhimurium.
Genetics. 1958 May;43(3):404-18
PMID: 17247766
-
A binding site for sulfate and its relation to sulfate transport into Salmonella typhimurium.
J Biol Chem. 1966 Sep 10;241(17):3962-9
PMID: 5331786
-
Classification of aminotransferase (C gene) mutants in the histidine operon.
J Mol Biol. 1966 Nov 14;21(2):335-55
PMID: 5339606
-
Regulation of sulfate transport in Salmonella typhimurium.
J Bacteriol. 1966 Jun;91(6):2275-80
PMID: 5329286
-
Selecting bacterial mutants by the penicillin method.
Science. 1960 Feb 26;131(3400):604-5
PMID: 13851300
-
Sulfate-binding protein from Salmonella typhimurium: physical properties.
Science. 1970 Jul 3;169(3940):59-61
PMID: 4911407
-
Nonsense mutants and polarity in the lac operon of Escherichia coli.
J Mol Biol. 1965 Nov;14(1):290-6
PMID: 5327654
-
The enzymic synthesis of L-cysteine in Escherichia coli and Salmonella typhimurium.
J Biol Chem. 1966 Nov 10;241(21):4955-65
PMID: 5332668
-
GENETIC STRUCTURE OF THE CYST REGION OF THE SALMONELLA GENOME.
Genetics. 1963 Aug;48:997-1009
PMID: 14054326
-
The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts.
J Biol Chem. 1965 Sep;240(9):3685-92
PMID: 4284300
-
Location of sulfate-binding protein in Salmonella typhimurium.
J Bacteriol. 1968 Oct;96(4):1049-54
PMID: 4971882
-
Cysteine Mutants of Salmonella Typhimurium.
Genetics. 1962 Nov;47(11):1617-27
PMID: 17248139
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Crystallization of a sulfate-binding protein (permease) from Salmonella typhimurium.
Science. 1967 Jun 23;156(3782):1627-8
PMID: 5337735
-
Positive control of sulphate reduction in Escherichia coli. The nature of the pleiotropic cysteineless mutants of E. coli K12.
Biochem J. 1968 Dec;110(3):597-602
PMID: 4882982
-
CHARACTERIZATION OF THE SULFITE AND HYDROXYLAMINE REDUCASES OF NEUROSPORA CRASSA.
J Biol Chem. 1965 Jun;240:2705-11
PMID: 14313747
-
Current linkage map of Salmonella typhimurium.
Bacteriol Rev. 1970 Jun;34(2):176-93
PMID: 4918632
-
Correlation between mutation type and the production of cross-reacting material in mutants of the A gene of the histidine operon in Salmonella typhimurium.
J Bacteriol. 1968 Feb;95(2):507-19
PMID: 4966548