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Role of chaperones and ATP synthase in DNA gyrase reactivation in Escherichia coli stationary-phase cells after nutrient addition.

Gutiérrez-Estrada A, Ramírez-Santos J, Gómez-Eichelmann Mdel C - Springerplus (2014)

Bottom Line: In these cells, the reactivation of DNA gyrase, which is a DNA topoisomerase type IIA enzyme, is responsible for the recovery of DNA supercoiling.Glucose addition to SP cells induced a slow recovery of DNA supercoiling, whereas resveratrol, which is an inhibitor of ATP synthase, inhibited the enzyme reactivation.These results suggest that DNA gyrase, which is an ATP-dependent enzyme, remains soluble in SP cells, and that its reactivation occurs primarily due to a rapid increase in the cellular ATP concentration.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology and Biotechnology, Institute of Biomedical Research, National Autonomous University of México, P.O. Box 70228, México City, 04510 México.

ABSTRACT
Escherichia coli stationary-phase (SP) cells contain relaxed DNA molecules and recover DNA supercoiling once nutrients become available. In these cells, the reactivation of DNA gyrase, which is a DNA topoisomerase type IIA enzyme, is responsible for the recovery of DNA supercoiling. The results presented in this study show that DNA gyrase reactivation does not require cellular chaperones or polyphosphate. Glucose addition to SP cells induced a slow recovery of DNA supercoiling, whereas resveratrol, which is an inhibitor of ATP synthase, inhibited the enzyme reactivation. These results suggest that DNA gyrase, which is an ATP-dependent enzyme, remains soluble in SP cells, and that its reactivation occurs primarily due to a rapid increase in the cellular ATP concentration.

No MeSH data available.


Related in: MedlinePlus

DNA gyrase reactivation in stationary-phase cells with low levels of polyphosphate, polyP. Cells grown in LB-MOPS media at 37°C. The strains used were BW25113 (wt) and BW25113 Δppk, bearing the reporter plasmid pMS01. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS medium. 1) Exponentially growing cells, 2) 48 hr stationary-phase cells, 3) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 30 sec, and 4) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 1 min. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.
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Fig2: DNA gyrase reactivation in stationary-phase cells with low levels of polyphosphate, polyP. Cells grown in LB-MOPS media at 37°C. The strains used were BW25113 (wt) and BW25113 Δppk, bearing the reporter plasmid pMS01. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS medium. 1) Exponentially growing cells, 2) 48 hr stationary-phase cells, 3) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 30 sec, and 4) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 1 min. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.

Mentions: SP wild-type and ppk cells grown in LB-MOPS at 37°C recover the SC level 30 sec-1 min after the addition of fresh medium; this recovery time is the shortest time reported using LB-MOPS medium (Reyes-Domínguez et al. 2003) (Figure 2). The results obtained with the E. coli mutant deficient in the enzyme involved in polyP synthesis suggest that this polymer is not involved in the fast reactivation of the DNA gyrase activity in starved cells after nutrient addition.Figure 2


Role of chaperones and ATP synthase in DNA gyrase reactivation in Escherichia coli stationary-phase cells after nutrient addition.

Gutiérrez-Estrada A, Ramírez-Santos J, Gómez-Eichelmann Mdel C - Springerplus (2014)

DNA gyrase reactivation in stationary-phase cells with low levels of polyphosphate, polyP. Cells grown in LB-MOPS media at 37°C. The strains used were BW25113 (wt) and BW25113 Δppk, bearing the reporter plasmid pMS01. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS medium. 1) Exponentially growing cells, 2) 48 hr stationary-phase cells, 3) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 30 sec, and 4) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 1 min. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4230433&req=5

Fig2: DNA gyrase reactivation in stationary-phase cells with low levels of polyphosphate, polyP. Cells grown in LB-MOPS media at 37°C. The strains used were BW25113 (wt) and BW25113 Δppk, bearing the reporter plasmid pMS01. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS medium. 1) Exponentially growing cells, 2) 48 hr stationary-phase cells, 3) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 30 sec, and 4) stationary-phase cells diluted 1:10 in LB-MOPS and incubated for 1 min. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.
Mentions: SP wild-type and ppk cells grown in LB-MOPS at 37°C recover the SC level 30 sec-1 min after the addition of fresh medium; this recovery time is the shortest time reported using LB-MOPS medium (Reyes-Domínguez et al. 2003) (Figure 2). The results obtained with the E. coli mutant deficient in the enzyme involved in polyP synthesis suggest that this polymer is not involved in the fast reactivation of the DNA gyrase activity in starved cells after nutrient addition.Figure 2

Bottom Line: In these cells, the reactivation of DNA gyrase, which is a DNA topoisomerase type IIA enzyme, is responsible for the recovery of DNA supercoiling.Glucose addition to SP cells induced a slow recovery of DNA supercoiling, whereas resveratrol, which is an inhibitor of ATP synthase, inhibited the enzyme reactivation.These results suggest that DNA gyrase, which is an ATP-dependent enzyme, remains soluble in SP cells, and that its reactivation occurs primarily due to a rapid increase in the cellular ATP concentration.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biology and Biotechnology, Institute of Biomedical Research, National Autonomous University of México, P.O. Box 70228, México City, 04510 México.

ABSTRACT
Escherichia coli stationary-phase (SP) cells contain relaxed DNA molecules and recover DNA supercoiling once nutrients become available. In these cells, the reactivation of DNA gyrase, which is a DNA topoisomerase type IIA enzyme, is responsible for the recovery of DNA supercoiling. The results presented in this study show that DNA gyrase reactivation does not require cellular chaperones or polyphosphate. Glucose addition to SP cells induced a slow recovery of DNA supercoiling, whereas resveratrol, which is an inhibitor of ATP synthase, inhibited the enzyme reactivation. These results suggest that DNA gyrase, which is an ATP-dependent enzyme, remains soluble in SP cells, and that its reactivation occurs primarily due to a rapid increase in the cellular ATP concentration.

No MeSH data available.


Related in: MedlinePlus