Intranuclear distribution, function and fate of glutathione and glutathione-S-conjugate in living rat hepatocytes studied by fluorescence microscopy

Bellomo, Giorgio; Palladini, G.; Vairetti, Mariapia

doi:10.1002/(sici)1097-0029(19970215)36:4<243::aid-jemt3>3.0.co;2-n

Cited by 51 publications

(27 citation statements)

References 53 publications

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“…Various studies have demonstrated that the nucleus is more reduced than the cytosol (15mM GSH vs. 11 mM, respectively) (Bellomo et al, 1997;Schafer & Buettner, 2001;Soboll et al, 1995). An important number of nuclear proteins, including transcription factors, require a reduced environment to bind to DNA.…”

Section: Modifications Of Nuclear Proteins Along the Cell Cyclesupporting

confidence: 90%

“…The importance of the GSH in cell proliferation could be extrapolated to the safeguarding of the nuclear architecture, providing in that way a proper milieu for the DNA replication. Our results could provide support to the hypothesis of Bellomo et al (Bellomo et al, 1997) that reduced nuclear glutathione may modulate the structural organization of chromatin. It is tempting to speculate that the high nuclear GSH level we observed before (late G1 phase) and at the onset of cell proliferation (S phase) could provide the redox environment that stimulates chromatin decompaction by reducing disulfide bonds.…”

Section: Pcna As a Possible Redox Sensor In The Onset Of Dna Synthesismentioning

confidence: 96%

“…The role of ATP-dependent mechanisms in maintaining the nuclear/cytoplasmic GSH concentration in hepatocytes was demonstrated by Bellomo and co-workers (Bellomo et al, 1997). After 20 min of incubation with the uncoupler protonophore carbonyl cyanide mchlorophenylhydrazone (CCCP) the nuclear/cytoplasmic GSH gradient disappeared, but the total GSH content remain unchanged.…”

Section: Atp Dependent Sequestration Of the Gsh To The Nucleusmentioning

confidence: 99%

See 2 more Smart Citations

The Nuclear Compartmentation of Glutathione: Effect on Cell Cycle Progression

Marković¹,

Mora²,

Gimeno³

et al. 2011

Selected Topics in DNA Repair

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Section: Modifications Of Nuclear Proteins Along the Cell Cyclesupporting

confidence: 90%

Section: Pcna As a Possible Redox Sensor In The Onset Of Dna Synthesismentioning

confidence: 96%

Section: Atp Dependent Sequestration Of the Gsh To The Nucleusmentioning

confidence: 99%

See 1 more Smart Citation

The Nuclear Compartmentation of Glutathione: Effect on Cell Cycle Progression

Marković¹,

Mora²,

Gimeno³

et al. 2011

Selected Topics in DNA Repair

View full text Add to dashboard Cite

“…The role of ATP-dependent mechanisms in maintaining the nuclear/cytoplasmic GSH concentration was studied by Bellomo et al (13,22) using culture hepatocytes pretreated with the uncoupler protonophore CCCP. After 20 min of incubation the nuclear/cytoplasmic GSH gradient disappeared, but the total GSH content remain unchanged.…”

Section: Discussionmentioning

confidence: 99%

Glutathione Is Recruited into the Nucleus in Early Phases of Cell Proliferation

Marković¹,

Borrás²,

Ortega

et al. 2007

Journal of Biological Chemistry

168

156

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We have studied the possible correlation between nuclear glutathione distribution and the progression of the cell cycle. The former was studied by confocal microscopy using 5-chloromethyl fluorescein diacetate and the latter by flow cytometry and protein expression of Id2 and p107. In proliferating cells, when 41% of them were in the S؉G 2 /M phase of the cell cycle GSH was located mainly in the nucleus. When cells reached confluence (G 0 /G 1 ) GSH was localized in the cytoplasm with a perinuclear distribution. The nucleus/cytoplasm fluorescence ratio for GSH reached a maximal mean value of 4.2 ؎ 0.8 at 6 h after cell plating. A ratio higher than 2 was maintained during exponential cell growth. In the G 0 /G 1 phase of the cell cycle, the nucleus/cytoplasm GSH ratio decreased to values close to 1. We report here that cells concentrate GSH in the nucleus in the early phases of cell growth, when most of the cells are in an active division phase, and that GSH redistributes uniformly between the nucleus and the cytoplasm when cells reach confluence.Glutathione (GSH) is the most abundant non-protein thiol in mammalian cells and performs many physiological functions (1). We have reported that cellular glutathione decreases in apoptosis (2).Although the role of nuclear GSH in the synthesis of DNA (3) and in protection against oxidative damage or ionizing radiation (4) is well established, little is known about the concentration of GSH in the nucleus and its regulation. This is due to two main factors. The first is methodological: it is impossible to determine the nuclear concentration of GSH using standard cell fractionation and analytical approaches (for a review see Söderdahl et al. (5). In view of this problem, we used confocal microscopy.The second factor is that most, if not all, of the reports share the common view of nuclear GSH distribution in a static situation. Cells are usually studied under steady state conditions i.e. when they are confluent (G 0 /G 1 phase of the cell cycle). The nucleus changes dramatically during the different phases of the cell cycle. Thus, studies addressed to determining the nuclear GSH distribution must take cell cycle physiology into account. To our knowledge there is a lack of information about the cellular distribution of glutathione during the different phases of the cell cycle and the possible correlation between cellular growth and nuclear GSH levels. We report here that GSH concentrates in the nucleus in the early phases of cell growth, when most of the cells are in an active division phase, and it redistributes uniformly between nucleus and cytoplasm when cells reach confluence. Nuclear Bcl-2 may be responsible for this change, as its expression changes in parallel with glutathione levels in nuclei. EXPERIMENTAL PROCEDURES Cell Culture3T3 fibroblasts were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum and antibiotics (25 units/ml penicillin, 25 g/ml streptomycin, and 0.3 g/ml amphotericin B) in 5% CO 2 in air at 37°C in 25 or 75 cm 2 f...

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“…It is interesting to point out that GSH distribution inside yeast cells is still not clear. Although it has been demonstrated already that GSH is distributed inside the mammalian nucleus at concentrations as high as in the cytoplasm (32), its presence inside the yeast nucleus is not clear, neither is the distribution of glutarredoxin isoforms inside yeast cells clear thus far. A mammalian nuclear isoform was described already (33,34) whereas information on yeast nuclear isoforms is still lacking.…”

Section: Discussionmentioning

confidence: 99%