Limits...
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

Effect of glucose as carbon source on DNA gyrase reactivation in MC4100 stationary-phase cells. Cells were grown in LB-MOPS at 37°C. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS or 1:30 in a 0.86% NaCl solution with or without 0.4% glucose. 1) Exponentially growing cells; 2) 48 hr stationary-phase cells; 3) and 4) stationary-phase cells diluted in LB-MOPS and incubated for 3 or 5 min, respectively; 5) and 6) stationary-phase cells diluted in 0.86% NaCl and incubated for 3 or 5 min, respectively; 7) and 8) stationary-phase cells diluted in 0.86% NaCl-0.4% glucose and incubated for 3 or 5 min, respectively. 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
getmorefigures.php?uid=PMC4230433&req=5

Fig3: Effect of glucose as carbon source on DNA gyrase reactivation in MC4100 stationary-phase cells. Cells were grown in LB-MOPS at 37°C. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS or 1:30 in a 0.86% NaCl solution with or without 0.4% glucose. 1) Exponentially growing cells; 2) 48 hr stationary-phase cells; 3) and 4) stationary-phase cells diluted in LB-MOPS and incubated for 3 or 5 min, respectively; 5) and 6) stationary-phase cells diluted in 0.86% NaCl and incubated for 3 or 5 min, respectively; 7) and 8) stationary-phase cells diluted in 0.86% NaCl-0.4% glucose and incubated for 3 or 5 min, respectively. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.

Mentions: As shown in Figure 3, SP cells recover DNA SC 3 min after dilution in LB-MOPS, whereas plasmid DNA remains relaxed in cells 3 or 5 min after dilution in 0.86% NaCl. However, cells diluted in 0.86% NaCl and 0.4% glucose partially recover the DNA SC level after 3 min of incubation and completely recover after 5 min. A plausible explanation is that after SP cell dilution in NaCl-glucose, the activation of genes specific for glucose, which is the added carbon source, induces energy production, e.g., ATP.Figure 3


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)

Effect of glucose as carbon source on DNA gyrase reactivation in MC4100 stationary-phase cells. Cells were grown in LB-MOPS at 37°C. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS or 1:30 in a 0.86% NaCl solution with or without 0.4% glucose. 1) Exponentially growing cells; 2) 48 hr stationary-phase cells; 3) and 4) stationary-phase cells diluted in LB-MOPS and incubated for 3 or 5 min, respectively; 5) and 6) stationary-phase cells diluted in 0.86% NaCl and incubated for 3 or 5 min, respectively; 7) and 8) stationary-phase cells diluted in 0.86% NaCl-0.4% glucose and incubated for 3 or 5 min, respectively. 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

Fig3: Effect of glucose as carbon source on DNA gyrase reactivation in MC4100 stationary-phase cells. Cells were grown in LB-MOPS at 37°C. To induce the recovery of the DNA SC level in stationary-phase cells, cell cultures were diluted 1:10 in pre-warmed LB-MOPS or 1:30 in a 0.86% NaCl solution with or without 0.4% glucose. 1) Exponentially growing cells; 2) 48 hr stationary-phase cells; 3) and 4) stationary-phase cells diluted in LB-MOPS and incubated for 3 or 5 min, respectively; 5) and 6) stationary-phase cells diluted in 0.86% NaCl and incubated for 3 or 5 min, respectively; 7) and 8) stationary-phase cells diluted in 0.86% NaCl-0.4% glucose and incubated for 3 or 5 min, respectively. Plasmid topoisomers were isolated and separated as described in Figure 1. Similar results were obtained in at least three independent experiments.
Mentions: As shown in Figure 3, SP cells recover DNA SC 3 min after dilution in LB-MOPS, whereas plasmid DNA remains relaxed in cells 3 or 5 min after dilution in 0.86% NaCl. However, cells diluted in 0.86% NaCl and 0.4% glucose partially recover the DNA SC level after 3 min of incubation and completely recover after 5 min. A plausible explanation is that after SP cell dilution in NaCl-glucose, the activation of genes specific for glucose, which is the added carbon source, induces energy production, e.g., ATP.Figure 3

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