Cation Transport Alteration Associated with Plasmid-Determined Resistance to Cadmium in
            <i>Staphylococcus aureus</i>

Weiss, Alison A.; Silver, Simón; Kinscherf, Thomas G.

doi:10.1128/aac.14.6.856

Cited by 75 publications

(49 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Cd'+ resistance in Gram-negative bacteria such as S. aureus, Bacillus subtilis and Lactobacillus plantarum is thought to involve a plasmid-coded Cd2+ efflux system or chromosomallydetermined reduced Cd2+ uptake (Weiss et a/., 1978;Laddaga et al, 1985b;Archibald & Duong, 1984). There is no clear evidence from the studies described here that Cd'+ resistance in the Gram-negative bacterium P. putida involves a Cd'+ efflux system.…”

Section: Discussionmentioning

confidence: 99%

Cadmium Resistance in Pseudomonas putida: Growth and Uptake of Cadmium

Higham¹,

Sadler²,

Scawen³

1985

Microbiology

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Cadmium Resistance in Pseudomonas putida: Growth and Uptake of Cadmium

Higham¹,

Sadler²,

Scawen³

1985

Microbiology

View full text Add to dashboard Cite

“…In S. aureus, Cd2' enters the cell by the Mn2' active transport system (4,6), but cells that have the cadA gene have lower net accumulation. The CadA-efflux system is sensitive to metabolic inhibitors such as uncouplers but is not affected by agents that eliminate the membrane potential (5).…”

mentioning

confidence: 99%

“…resistance function coded by the cadA determinant results from decreased intracellular accumulation of Cd2+ (4), mediated by an energy-dependent efflux mechanism (5).…”

mentioning

confidence: 99%

Cadmium resistance from Staphylococcus aureus plasmid pI258 cadA gene results from a cadmium-efflux ATPase.

Nucifora

Chu

Misra

et al. 1989

Proc. Natl. Acad. Sci. U.S.A.

330

244

View full text Add to dashboard Cite

Cadmium resistance specified by the cadA determinant of Staphylococcus aureus plasmid p1258 results from the functioning of a cadmium-efflux system. In the nucleotide sequence of the DNA fragment containing the cadA determinant, two open reading frames were identified. The larger one, corresponding to a predicted polypeptide of 727 amino acid residues, is necessary and sufficient for expression of cadmium resistance. (no. 54 Kodak Wratten filter) at 37TC with shaking, harvested by centrifugation at 6800 X g for 7 min, and suspended in medium at a density of25 mg (dry weight)/ml. The final cell density in transport assays was 0.5 mg (dry weight)/ml. Transport assays were at 37°C in broth with aeration by shaking. Samples were filtered through 0.45-,um pore-diameter filters (Bio-Rad) and rinsed twice with 5 ml ofNaCl solution (9 g/liter). Radioactive samples were assayed in a liquid scintillation spectrometer (Packard Instrument).' DNA Cloning and Sequencing. For construction of plasmids and phage M13 derivatives containing the cadA determinant, a derivative of pRALl (1, 9) was used. The 3.5-kilobase (kb) Bgl II-Xba I DNA fragment, previously identified by Novick et al.(2) as containing the cadA Cd2+-resistance determinant, was cloned into phage mTMO10 (10) in both orientations. Nested deletions for sequencing were made by using BAL-31 exonuclease (10). The 3.5-kb fragment was also cloned into vector pSK265 (7) (for expression in B. subtilis). Two recombinant plasmids, pGN114 and pGN115, containing the entire 3.5-kb DNA fragment in opposite orientations, were generated.Plasmids pGN116-1 and pGN116-2 [containing the intact larger ORF (ORF2, cadA) cloned in both orientations] were obtained by ligating into the vector plasmid pSK265 a DNA fragment generated by BAL-31 deletion and lacking the first 920 base pairs (bp) of the Bgl II-Xba I fragment shown in Fig.

show abstract

“…In the present report, the induction pattern of the cadA operon by various toxic heavy metal cations is described. The cadA cadmium resistance system was previously thought to function constitutively (8,12,20,22) stop sequence, were the gifts of R. P. Novick and were used to construct cadA-blaZ fusions. Cell growth on 2XNY medium, the use of the antibiotics ampicillin (100 ,ug/ml), erythromycin (10 ,ug/ml), and chloramphenicol (5 ,ug/ml), and measurements of growth by turbidity were as described in the accompanying report (24).…”

mentioning

confidence: 99%

“…In the present report, the induction pattern of the cadA operon by various toxic heavy metal cations is described. The cadA cadmium resistance system was previously thought to function constitutively (8,12,20,22), although this hypothesis was not directly tested. As the results of our new research have shown, the inability to demonstrate regulated control of cadA was largely due to the use of Cd2' both as an inducer and as a toxic challenge compound.…”

mentioning

confidence: 99%

Regulation of the cadA cadmium resistance determinant of Staphylococcus aureus plasmid pI258

1991

View full text Add to dashboard Cite

Regulation of the cadA cadmium and zinc resistance determinant of Staphylococcus aureus plasmid p1258 was demonstrated by using gene fusions and direct measurements of transcription. In growth experiments, cells harboring the intact cadA operon were induced with different cations and challenged by an inhibitory concentration of ZnC12, a substrate of the CadA resistance system. Uninduced cells did not grow for 8 h after Zn2+ addition, whereas induced cells grew in the presence Zn2+. Cd2" was a strong inducer, and Bi3+ and Pb2+ also induced well; Co2, and Zn2+ were weak inducers. A translational (-lactamase fusion to the cadA gene showed the same induction specificity as that seen with growth experiments with the intact cadA operon. A short j-lactamase transcriptional fusion to the cadC gene also showed the same pattern of induction, establishing that the cadC gene was not involved in regulation. In Northern (RNA) blot hybridization experiments, a cadmium-inducible, 2.6-kb, operon-length transcript was detected. Primer extension experiments determined that Cd2+-inducible transcription of the cad4 operon begins at nucleotides 676 and 677 of the published sequence (G. Nucifora, L. Chu, T. K. Misra, and S. Silver, Proc. Natl. Acad. Sci. USA 86: [3544][3545][3546][3547][3548] 1989).The cadA cadmium resistance determinant of Staphylococcus aureus plasmid pI258 contains two open reading frames (8). The protein products of the cadA efflux ATPase gene (727 amino acids) and of the shorter cadC gene (122 amino acids) have been identified, and the need for both genes for cadmium and zinc resistance has been established in the accompanying paper (24). In the present report, the induction pattern of the cadA operon by various toxic heavy metal cations is described. The cadA cadmium resistance system was previously thought to function constitutively (8,12,20,22) stop sequence, were the gifts of R. P. Novick and were used to construct cadA-blaZ fusions. Cell growth on 2XNY medium, the use of the antibiotics ampicillin (100 ,ug/ml), erythromycin (10 ,ug/ml), and chloramphenicol (5 ,ug/ml), and measurements of growth by turbidity were as described in the accompanying report (24). Cell mass as dry weight was calculated from a curve of dry weight versus turbidity (6).DNA procedures. The manipulation of DNA with enzymes was as described by Sambrook et al. (13). The structure of each fusion was confirmed by nucleotide sequencing by the dideoxynucleotide method (15).Materials. Cadmium chloride, cobalt dichloride, zinc chloride, manganese chloride, lead chloride, and potassium bismuth tartrate were used as cation salts. 5-Bromo-4-chloro-3-indolyl-p-D-galactopyranoside (X-Gal) and isopropyl-p-D-thiogalactopyranoside (IPTG)

show abstract

Cation Transport Alteration Associated with Plasmid-Determined Resistance to Cadmium in Staphylococcus aureus

Cited by 75 publications

References 30 publications

Cadmium Resistance in Pseudomonas putida: Growth and Uptake of Cadmium

Cadmium Resistance in Pseudomonas putida: Growth and Uptake of Cadmium

Cadmium resistance from Staphylococcus aureus plasmid pI258 cadA gene results from a cadmium-efflux ATPase.

Regulation of the cadA cadmium resistance determinant of Staphylococcus aureus plasmid pI258

Contact Info

Product

Resources

About