**5.2 Nuclear synthesis of glutathione**

Glutamate cysteine ligase (GCL) and GSH synthetase activities have been reported in nuclei, and a portion of 4-8% of cell GSH synthetic activity is considered capable of maintaining nuclear GSH levels (Ho and Guenthner, 1997). However, we could not find GCL expression in nuclei of 3T3 fibroblasts. In addition as previously reported BSO, a specific inhibitor of GCL was unable of decreasing nuclear glutathione levels. Thus, at least under our experimental conditions, the possible "de novo" synthesis of nuclear glutathione seems improbable in 3T3 fibroblasts.

#### **5.3 GSH enters nucleus via nuclear pores**

The nuclear pore complex is the biochemical machinery that controls the molecular traffic across the nuclear envelope (Feldherr & Akin, 1990; Nigg, 1997). Ions and small hydrophilic molecules, like glutathione, are considered to move by free and fast diffusion across the nuclear pore (Ribbeck & Gorlich, 2001); nevertheless ion gradients and transnuclear ATPdependent membrane potential have also been reported (Nigg, 1997). In a series of creative experiments published in early 1990ies, Feldherr CM and Akin D (Feldherr & Akin, 1990; Feldherr & Akin, 1993), shown that permeability of nuclear envelope and nuclear transport were higher in proliferating than in quiescent cells. Reported seven fold reduce in the nuclear transport capacity was induced by the alterations in the characteristics of the pores and not by the changes within the cytoplasm, specifically, the decrease in ATP concentration. One pore forming protein that has been brought into the connection to nuclear glutathione content is Bcl-2. Voehringer and colleagues (Voehringer et al., 1998) showed that over-expression of Bcl-2 recruits GSH to the nucleus. The presence of this protein at the nuclear envelope was demonstrated (Krajewski et al., 1993) and the association with the nuclear pore complexes was suggested. Moreover, Zimmermann et al. (Zimmermann et al., 2007) demonstrated that GSH binds to Bcl-2 in mitochondria, providing a molecular basis for its antioxidant function.

A clear picture emerges showing that the presence of a reduced nuclear environment, probably provided by glutathione, glutaredoxin and thioredoxin mainly, is of paramount importance in the physiology of cell cycle, underscoring the role of oxidative stress in cell proliferation.

#### **6. References**

284 Selected Topics in DNA Repair

nuclear GSH levels, and not total cellular glutathione levels, that specifically correlate with cellular proliferation. Glutathione is considered essential for survival in mammary cells and other eukaryotic cells, but not prokaryotic cells. However, although a number of important functions have been attributed to GSH, its outstanding role in nucleated, but not in prokaryotic cells, remains unknown. Our results underscore the important role of nuclear glutathione in cell physiology and suggest that manipulation of nuclear GSH levels could be

**5. The occurrence of the glutathione in the nucleus; active transport, de novo** 

How GSH enters the nucleus and how it is regulated during the different phases of the cell cycle is still a matter of debate. The regulation of such interactions is also unclear. According to Smith and colleagues (Smith et al., 1996), the possible biochemical mechanisms responsible for the turnover of nuclear GSH are the following: 1.GSH may be taken up from the cytoplasm into the nuclei either passively or through energy-dependent processes 2.GSH may be synthesized *de novo* in the nucleus by the enzymes glutamate cysteine ligase

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. This study was questioned by Briviba et al (Briviba et al., 1993) because of the use of monochlorobimane (BmCl). Despite its high specificity for glutathione this fluorochrome was found to be of no value in the study of cellular GSH distribution; once GSH-BmCl conjugate is formed it demonstrates an increased tendency of nuclear compartmentalization. Indeed, in our study using CMFDA, we have not found an ATP-dependent mechanism of nuclear GSH compartmentalization in 3T3 fibroblasts. Ho and Guenthner, 1997 using nuclear fractions concluded that GSH is taken up by the nucleus by passive diffusion and no evidence for an ATP-dependent mechanism for

Glutamate cysteine ligase (GCL) and GSH synthetase activities have been reported in nuclei, and a portion of 4-8% of cell GSH synthetic activity is considered capable of maintaining nuclear GSH levels (Ho and Guenthner, 1997). However, we could not find GCL expression in nuclei of 3T3 fibroblasts. In addition as previously reported BSO, a specific inhibitor of GCL was unable of decreasing nuclear glutathione levels. Thus, at least under our experimental conditions, the possible "de novo" synthesis of nuclear glutathione seems

The nuclear pore complex is the biochemical machinery that controls the molecular traffic across the nuclear envelope (Feldherr & Akin, 1990; Nigg, 1997). Ions and small hydrophilic

of paramount importance during development and cancer.

and GSH synthetase 3.GSH may function to transport -glu-cys-cys.

**5.1 ATP dependent sequestration of the GSH to the nucleus** 

**synthesis, diffusion or something else** 

GSH concentration was observed.

improbable in 3T3 fibroblasts.

**5.2 Nuclear synthesis of glutathione** 

**5.3 GSH enters nucleus via nuclear pores** 


The Nuclear Compartmentation of Glutathione: Effect on Cell Cycle Progression 287

Dijkwel, P.A. & Wenink, P.W. (1986). Structural integrity of the nuclear matrix: differential

Dimitrova, D.S. & Gilbert, D.M. (2000). Temporally coordinated assembly and disassembly

Dimitrova, D.S. & Berezney, R. (2002). The spatio-temporal organization of DNA replication

Esposito, F., Russo, T. & Cimino, F. (2002). Generation of prooxidant conditions in intact

Fratelli, M,; Goodwin, L.O.; Ørom, U.A., Lombardi, S., Tonelli, R., Mengozzi, M. & Ghezzi,

Feldherr, C.M. & Akin, D. (1993). Regulation of nuclear transport in proliferating and

Fujii, H.; Li, S.H.; Szmitko, P.E.; Fedak, P.W. Verma, S. (2006). C-reactive protein alters

García-Cao, M.; O'Sullivan, R.; Peters, A.H.; Jenuwein, T. Blasco, M.A. (2004). Epigenetic

Giustarini, D.; Rossi, R.; Milzani, A.; Colombo, R. Dalle-Donne, I. (2004). S-glutathionylation:

Grant, C.M.; MacIver, F.H. Dawes, I. W. (1996). Glutathione is an essential metabolite

Green, R.M.; Graham, M.; O'Donovan, M.R., Chipman, J.K. & Hodges, N.J. (2006).

*molecular medicine*, 8, 2, (April 2004), pp. 201-212, ISSN 1582-1838.

*Current Genetics*, 29, 6, (May 1996), pp. 511-515, ISSN 0172-8083.

*America,* 102, 39, (September 2005) pp. 13998-14003, ISSN 0027-8424. Feldherr, C.M. & Akin, D. (1990). The permeability of the nuclear envelope in dividing and

ISSN 0449-3060.

0027-8424.

0021-9525.

0014-4827.

1061-4036.

2482 ISSN 1079-5642.

383-390, ISSN 0267-8357.

pp. 53-67, ISSN 0021-9533.

(October 2000), pp. 686-694, ISSN 1465-7392.

*Enzymology,* 352, pp. 258-268, ISSN 0076-6879.

mammary carcinoma cells. *Radiation Research*, 114, 2, (November 1988), pp. 215-224,

effects of thiol agents and metal chelators*. Journal of cell science*, 84, (August 1986)

of replication factories in the absence of DNA synthesis. *Nature cell biology*, 2, 10,

sites is identical in primary, immortalized and transformed mammalian cells. *Journal of cell science,* 115, Pt 21, (November 2002) pp. 4037-4051, ISSN 0021-9533. Ernst, V.; Levin, D.H. London, I.M. (1978.) Inhibition of protein synthesis initiation by

oxidized glutathione: activation of a protein kinase that phosphorylates the alpha subunit of eukaryotic initiation factor 2. *Proceedings of the National Academy of Sciences of the United States of America*, 75, 9, (September 1978), pp. 4110-4114, ISSN

cells to induce modifications of cell cycle regulatory proteins*. Methods in* 

P. (2005). Gene expression profiling reveals a signaling role of glutathione in redox regulation. *Proccedings of the National Academy of Science of the United States of* 

nondividing cell cultures. *The Journal of cell biology,* 111, 1 (July 1990) pp. 1-8 ISSN

quiescent cells. *Experimental cell research*, 205, 1, (March 1993), pp. 179-186, ISSN

antioxidant defenses and promotes apoptosis in endothelial progenitor cells. *Arteriosclerosis, thrombosis, and vascular biology,* 26, 11, (November 2006), pp. 2476-

regulation of telomere length in mammalian cells by the Suv39h1 and Suv39h2 histone methyltransferases. *Nature Genetics*, 36, 1, (January 2004), pp. 94-99, ISSN

from redox regulation of protein functions to human diseases. *Journal of cellular and* 

required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae.

Subcellular compartmentalization of glutathione: correlations with parameters of oxidative stress related to genotoxicity*. Mutagenesis*, 21, 6, (Novembre 2006), pp.


Boyland, E. & Chasseaud L.F. (1969). Glutathione S-aralkyltransferase. *The Biochemical* 

Britten, R.A.; Green, J.A.; Broughton, C.; Browning, P.G.; White, R. & Warenius H. M. (1991).

Briviba, K.; Fraser, G.; Sies, H. & Ketterer, B. (1993). Distribution of the monochlorobimane-

*Biochemical Journal*, 294, Pt 3, (September 1993), pp. 631-633, ISSN 0264-6021. Brown, K.E.; Meleah Mathahs, M.; Broadhurst, K.A.; Coleman, M.C.; Ridnour, L.A.;

Burchill, B.R.; Oliver, J.M.; Pearson, C.B.; Leinbach, E.D. Berlin, R.D. (1978). Microtubule

Canela, A.; Vera, E.; Klatt, P. Blasco, M.A. (2007). High-throughput telomere length

Carpenter, G.& Cohen, S. (1990). Epidermal growth factor*. The Journal of Biological Chemistry*,

Castillo, P.; Bogliolo, M. & Surralles J. (2011). Coordinated action of the Fanconi anemia and

Conour, J.E.; Graham, W.V. & Gaskins, H.R. (2004). A combined in vitro/bioinformatic

Dai, X.; Huang, C.; Bhusari, A., Sampathi, S.; Schubert, K. & Chai, W. (2010). Molecular steps

Dalle-Donne, I.; Rossi, R.; Giustarini, D.; Colombo, R. Milzani, A. (2007). S-

Davies, K.J. (1999). The broad spectrum of responses to oxidants in proliferating cells: a new

De Rey Pailhade, M. (1988). Sur un corps d'origine organique hydrogénant le soufre á froid.

Dethlefsen, L.A.; Lehman, C.M.; Biaglow, J.E. Peck, V.M. (1988). Toxic effects of acute

*Physiological Genomics*, 18, 2 (July 2004), pp. 196-205, ISSN 1094-8341. Cotgreave, I.A. Gerdes, R.G. (1998). Recent trends in glutathione biochemistry--

The relationship between nuclear glutathione levels and resistance to melphalan in human ovarian tumour cells*. Biochemical Pharmacology*, 41, 4, (February 1991) pp.

glutathione conjugate between nucleus and cytosol in isolated hepatocytes. *The* 

Schmidt, W.N. Spitz, D.R. (2007). Increased hepatic telomerase activity in a rat model of iron overload: a role for altered thiol redox state? *Free Radical Biology* 

dynamics and glutathione metabolism in phagocytizing human polymorphonuclear leukocytes. *Journal of Cellular Physiology*, 76, 2, (February 1978),

quantification by FISH and its application to human population studies. *Proceedings of the National Academy of Sciences of the United States of America,* 104, 13 (March

ataxia telangiectasia pathways in response to oxidative damage. *DNA Repair(Amst).* 

investigation of redox regulatory mechanisms governing cell cycle progression.

glutathione-protein interactions: a molecular link between oxidative stress and cell proliferation? *Biochemical and biophysical research communications*, 242, 1, (January

of G-overhang generation at human telomeres and its function in chromosome end protection. *The EMBO Journal,* 29, 16 (August 2010), pp. 2788-2801, ISSN 0261-4189.

glutathionylation in protein redox regulation. *Free Radical Biology Medicine*, 43, 6,

paradigm for oxidative stress. *IUBMB Life,* 48, 1, (July 1999), pp. 41-47, ISSN 1521-

glutathione depletion by buthionine sulfoximine and dimethylfumarate on murine

*Journal.* 115, 5, (Decembre 1969), pp. 985-991, ISSN 0264-6021.

*Medicine*, 42, 2, (January 2007), pp. 228-235, ISSN 0891- 5849.

647-649, ISSN 0006-2952.

pp. 439-447, ISSN 0021-9541.

2007), pp.5300-5305, ISSN 0027-8424.

1998), pp. 1-9, ISSN 0006-291X.

6543.

265, 14 (May 1990), pp. 7709-7712, ISSN 0021-9258.

10, 5 (May 2011), pp. 518-525, ISSN 1568-7864.

(September 2007), pp. 883-898, ISSN 0891-5849.

*C.R. Acad. Sci.* , 106, pp. 1683-1684, ISSN 1287-4620.

mammary carcinoma cells. *Radiation Research*, 114, 2, (November 1988), pp. 215-224, ISSN 0449-3060.


The Nuclear Compartmentation of Glutathione: Effect on Cell Cycle Progression 289

Jády, B.E.; Richard, P.; Bertrand, E. Kiss, T. (2006). Cell cycle-dependent recruitment of

Jang, J.H. &. Surh, Y.J. (2003). Potentiation of cellular antioxidant capacity by Bcl-2:

Kennedy BK, Barbie DA, Classon M, Dyson N, Harlow E. (2000). Nuclear organization of

Kim, N.W.; Piatyszek, M.A.; Prowse, K.R.; Harley, C.B.; West, M.D.; Ho, P.L.; Coviello,

Krajewski, S.; Tanaka, S.; Takayama, S.M.; Schibler, M.J.; Fenton, W. & Reed J.C. (1993).

Kosower, N.S. Kosower, E.M. (1978). The glutathione status of cells. *International Review* 

Koziel. J.E.; Fox, M.J.; Steding, C.E.; Sprouse, A.A. & Herbert, B.S. (2011). . Medical genetics

Lasorella, A.; Iavarone, A. & Israel, M.A. (1996). Id2 specifically alters regulation of the cell

Leonhardt, H.; Rahn, H.P.; Weinzierl, P.; Sporbert, A.; Cremer, T.; Zink, D. & Cardoso M.C.

Martensson, J.; Lai, J.C. & Meister, A. (1990). High-affinity transport of glutathione is part of

Meister, A. Anderson, M. E. (1983). Glutathione. *Annual Review of Biochemistry*, 52, (1983),

Menon, S.G.; Sarsour, E.H.; Spitz, D.R.; Higashikubo, R.; Sturm, M.; Zhang, H. Goswami,

*Cancer Research*, 53, 19, (October 1993), pp. 4701-4714, ISSN 0008-5472. Kornberg, A. (1969). Active center of DNA polymerase. *Science*, 163, 874, (March 1969), pp.

17, 2, (February 2006), pp. 944-954, ISSN 1059-1524.

(September-October 2006), pp. 1865-1879, ISSN 1523-0864.

2003), pp. 1371-1379, ISSN 0006-2952.

1410-1418, ISSN 0036-8075.

pp. 2570-2578, ISSN 0270-7306.

pp. 7185-7189, ISSN 0027-8424.

pp. 711-760, ISSN 0066-4154.

Jones, D.P. (2006). Redefining oxidative stress. *Antioxidants* 

(Novembre 2000), pp. 2855-2568, ISSN 0890-9369.

(December 1994), pp. 2011-2015, ISSN 0036-8075.

*Cytology*, 54, (1978), pp. 109-160, ISNN 0074-7696.

*Biology*, 149, 2, (April 2000), pp. 271-280, ISSN 0021-9525.

(March, 2011), pp. 457-467 ISNN 1582-1838.

telomerase RNA and Cajal bodies to human telomeres. *Molecular Biology of the Cell*,

implications for its antiapoptotic function. *Biochemical Pharmacology*, 66, 8, (October

DNA replication in primary mammalian cells. *Genes and Development*, 14, 22,

G.M.; Wright, W.E.; Weinrich, S.L. Shay, J.W. (1994). Specific association of human telomerase activity with immortal cells and cancer. *Science*, 266, 5193,

Investigation of the subcellular distribution of the Bcl-2 oncoprotein: residence in the nuclear envelope, endoplasmic reticulum, and outer mitochondrial membranes.

and epigenetics of telomerase. *Journal of cellular and molecular medicine*, 15, 3,

cycle by tumor suppressor proteins. *Molecular and cellular biology,* 16, 6, (June 1996),

(2000). Dynamics of DNA replication factories in living cells. *The Journal of Cellular* 

a multicomponent system essential for mitochondrial function. *Proccedings of the National Academy of Sciences of the United States of America,* 87, 18, (September 1990),

P.C. (2003). Redox regulation of the G1 to S phase transition in the mouse embryo fibroblast cell cycle. *Cancer Research*, 63, 9, (May 2003), pp. 2109-2117 ISSN 1078-0432. Menon, S.G. Goswami, P.C. (2007). A redox cycle within the cell cycle: ring in the old with the new. *Oncogene*, 26, 8, (February 2007), pp. 1101-1109, ISSN 0950-9232. Michishita, E.; McCord, R.A.; Berbe,r E., Kioi, M., Padilla-Nash, H., Damian, M., Cheung, P.;

Kusumoto, R., Kawashara, T.L.; Barrett, JC.; Chang, H.Y.; Bohr, V.A.; Ried, T.; Gozani, O. & Chua, K.E. (2008). SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. *Nature*. 452, 7186, (March 2008), pp. 492-496, ISSN 0028- 0836.

 *Redox Signaling*, 8, 9-10,


Greenberg, J.T. Demple, B. (1986). Glutathione in Escherichia coli is dispensable for

Guo, Z.; Kozlov, S.; Lavin, M.F.; Person, M.D. & Paull T. (2010)ATM Activation by Oxidative

Halliwell, B. (2007). Oxidative stress and cancer: have we moved forward?. *Biochemical* 

Hansen, J.M.; Go, Y.M. & Jones, D.P. (2006). Nuclear and mitochondrial

Hayakawa, N.; Nozawa, K.; Ogawa, A.; Kato, N.; Yoshida, K.; Akamatsu, K.; Tsuchiya, M.;

Ho, Y.F. & Guenthner, T.M. (1997). Isolation of liver nuclei that retain functional trans-

Hoffman, A.; Spetner, L.M. & Burke, M. (2008). Ramifications of a redox switch within a

Holmgren, A. (1976). Hydrogen donor system for Escherichia coli ribonucleoside-

Hopkins, F. (1929). On glutathione: a reinvestigation. *Journal of Biological Chemistry*, 84,

Huang, Z.Z.; Chen, C.; Zeng, Z.; Yang, H., Oh, J.; Chen, L, & Lu, S.C. (2001). Mechanism and

Hug, N. & Lingner, J. (2006). Telomere length homeostasis. *Chromosoma,* 115, 6, (December

Hutter, D.E.; Till, B.G. & Greene, J.J. (1997). Redox state changes in density-dependent

Hwang, C., A.J. Sinskey, and H.F. Lodish, (1992). Oxidized redox state of glutathione in the

*Pharmacology and Toxicol*ogy 46, (XXX 2006), pp. 215-234. ISSN 0362-1642. Harley, C.B.; Futcher, A.B. Greider, C.W. (1990). Telomeres shorten during ageing of human fibroblasts. *Nature*, 345, 6274, (May 1990), pp. 458-460, ISSN 0028-0836. Harris, J.W. and Patt, H.M. (1969). Non-protein sulfhydryl content and cell-cycle dynamics

Stress. *Science*. 22, 330, (October, 2010), pp. 517-521 ISSN 0193-4511. Haendeler, J.; Hoffmann, J.; Diehl, J.F.; Vasa, M.; Spyridopo ulos, I.; Zeiher, A.M.

*Research*, 94, 6, (April 2004), pp. 768-775, ISSN 0009-7330.

*Journal*, 401, 1, (January 2007), pp. 1-11, ISSN 0264-6021.

1986), pp. 1026-1029, ISSN 0021-9193.

1999), pp. 11501-11507, ISSN 0006-2960.

(Novembre 1997), pp. 163-168, ISSN 1056-8719.

(August 2008), pp. 265-268, ISSN 0891-5849.

(1929), pp. 269-320, ISSN 0021-9258.

2006), pp. 413-25, ISSN 0009-5915.

ISSN 0014-4827.

ISSN 0027-8424.

438, ISSN 0014-4827.

0193-4511.

6638.

resistance to H2O2 and gamma radiation. *Journal of Bacteriology*, 168, 2, (November

Dimmeler, S. (2004). Antioxidants inhibit nuclear export of telomerase reverse transcriptase and delay replicative senescence of endothelial cells. *Circulation* 

compartmentalization of oxidative stress and redox signaling*. Annual Review of* 

of Ehrlich ascites tumor. *Experimental Cell Research*, 56, 1, (July 1969), pp. 134-141,

Nagasaka, A. Yoshida, S. (1999). Isothiazolone derivatives selectively inhibit telomerase from human and rat cancer cells in vitro. *Biochemistry*, 38, 35, (August

membrane transport. *Journal of Pharmacological and Toxicological Methods,* 38, 3,

normal cell: its absence in a cancer cell. *Free Radical Biology and Meicine*, 45, 3,

diphosphate reductase dependent upon glutathione. *Proceedings of the National Academy of Sciences of the United States of America*, 73, 7, (July 1976), pp. 2275-2279,

significance of increased glutathione level in human hepatocellular carcinoma and liver regeneration. *The Faseb Journal,* 15, 1, (January 2001), pp. 19-21, ISSN 0892-

regulation of proliferation. *Experimental Cell Research*, 232, 2, (May 1997), pp. 435-

endoplasmic reticulum. *Science,* 257, 5076, (September 1992), pp. 1496-1502, ISSN


The Nuclear Compartmentation of Glutathione: Effect on Cell Cycle Progression 291

Rodriguez, J.L.; Sandoval, J.; Serviddio, G.; Sastre, J.; Morante, M.; Perrelli, M.G.; Martínez-

Rodríguez-Manzaneque, M.T.; Ros, J.; Cabiscol, E.; Sorribas, A. Herrero, E. (1999). Grx5

Schafer, F.Q. Buettner, G.R. (2001). Redox environment of the cell as viewed through the

Sen, C.K. & Packer, L. (1996). Antioxidant and redox regulation of gene transcription. *The* 

Sharpless, N.E. DePinho, R. A. (2004). Telomeres, stem cells, senescence, and cancer. *Journal of Clinical Investigation*, 113, 2, (January 2004), pp. 160-168, ISSN 0021-9738. Sies, H. Cadenas, E. (1985). Oxidative stress: damage to intact cells and organs.

Sies, H. (1999). Glutathione and its role in cellular functions. *Free Radical Biology Medicine*, 27,

Smith, J.E. (1977). Elevated erythrocyte glutathione associatd with elevated substrate in

Smith, C.V.; Jones, D.P.; Guenthner, T.M.; Lash, L.H. & Lauterburg, B.H. (1996).

Soboll, S.; Gründel, S.; Harris, J.; Kolb-Bachofen. V.; Ketterer, B. & Sies, H. (1995). The

Söderdahl ,T.; Enoksson, M.; Lundberg, M.; Holmgren, A.; Ottersen, O.P.; Orrenius, S.;

Spector, D.; Labarre, J. Toledano, M.B. (2001). A genetic investigation of the essential role

Spyrou, G. & Holmgren, A. (1996). Deoxyribonucleoside triphosphate pools and growth of

Sun, Y. Oberley, L.W. (1996). Redox regulation of transcriptional activators. *Free Radical* 

*Communications*, 220, 1, (March 1996), pp. 42-46, ISSN 0006-291X.

*Biology Medicine*, 21, 3, (1996), pp. 335-348, ISSN 0891- 5849.

*Journal*, 17, 1, (January 2003), pp. 124-126, ISSN 0892-6638.

(March 2001), pp. 7011-7016, ISSN 0042-6989.

398, 3, (September 2006) pp. 431-437, ISSN 0264-6021.

*Medicine*, 30, 11, (June 2001), pp. 1191-212, ISSN 0891- 5849.

1152, (December 1985), pp. 617-631, ISSN 0962-8436.

9-10, (November 1999), pp. 916-921, ISSN 0891- 5849.

*FASEB Journal,* 10, 7, (May 1996) pp. 709-720, ISSN 0892-6638.

pp. 8180-8190, ISNN 0270- 7306.

1977), pp. 516-520, ISSN 006-3002.

ISSN 0041-008X.

ISSN 0264-6021.

Chantar, M.L.; Viña, J.; Viña, J.R.; Mato, J.M.; Avila, M.A.; Franco, L.; López-Rodas, G.& Torres, L. (2006). Id2 leaves the chromatin of the E2F4-p130-controlled c-myc promoter during hepatocyte priming for liver regeneration. *The Biochemical Journal*,

gluta redoxin plays a central role in protection against protein oxidative damage in Saccharomyces cerevisiae. *Molecular and Cellular Biology*, 19, 12, (December 1999),

redox state of the glutathione disulfide/glutathione couple. *Free Radical Biology* 

*Philosophical transactions of the Royal Society of London. Series B, Biological sciences*, 311,

high-and low-glutathione sheep. *Biochemica et Biophysica Acta,* 496, 2, (February

Compartmentalization of glutathione: implications for the study of toxicity and disease. *Toxicology and Applied Pharmacology,* 140, 1, (September 1996), pp. 1-12,

content of glutathione and glutathione S-transferases and the glutathione peroxidase activity in rat liver nuclei determined by a non-aqueous technique of cell fractionation. *The Biochemical Journal*, 311, Pt 3, (November 1995), pp. 889-894,

Bolcsfoldi, G. Cotgreave, I.A. (2003). Visualization of the compartmentalization of glutathione and protein-glutathione mixed disulfides in cultured cells. *Faseb* 

of glutathione: mutations in the proline biosynthesis pathway are the only suppressors of glutathione auxotrophy in yeast. *Journal of Biology Chemistry*, 276, 10,

glutathione-depleted 3T6 mouse fibroblasts. *Biochemical and Biophysical Research* 


Minamino, T.; Mitsialis, S.A. Kourembanas, S. (2001). Hypoxia extends the life span of

Miranda-Vizuete, A.; Rodríguez-Ariza, A.; Toribio, F.; Holmgren, A.; López-Barea, J.

Nigg, E.A. (1997). Nucleocytoplasmic transport: signals, mechanisms and regulation. *Nature*,

Nkabyo, Y.S.; Ziegler, T.R.; Gu, L.H.; Watson, W.H. Jones, D.P. (2002). Glutathione and

Norton, J.D. (2000). ID helix-loop-helix proteins in cell growth, differentiation and

Oberley, L.W.; Oberley, T.D. Buettner, G.R. (1981). Cell division in normal and

Oleinick, N.L.; Chiu, S.M.; Ramakrishnan, N. & Xue, L.Y. (1987). The formation,

Pani, G.; Colavitti, R.; Bedogni, B.; Anzevino, R.; Borrello, S. Galeotti, T. (2000). A redox

Post, G.B., Keller, DA.; Connor, K.A. & Menzel, D.B. (1983) Effects of culture conditions on

Reddy, N.M.; Kleeberger, S.R.; Bream, .JH.; Fallon, P.G.; Kensler, T.W.; Yamamoto, M. &

Reynaert, N.L.; van der Vliet, A.; Guala, A.S.; McGovern, T.; Hristova, M.; Pantano, C.;

Ribbeck, K. & Gorlich, D. (2001). Kinetic analysis of translocation through nuclear pore complexes. *The EMBO Journal*, 20, 6, (March 2001): pp. 1320-1330, ISSN 0261-4189.

*Biology*, 21, 10, (May 2001), pp. 3336-3342, ISSN 0270-7306.

32, (August 1996), pp. 19099-19103, ISSN 0042-6989.

386, 6627, (April 1997), pp. 779-787, ISSN 0028- 0836.

*Hypotheses*, 7, 1, (January 1981), pp. 21-42, ISSN 0306-9877.

*America,* 77, 12 (December 1980), pp. 127315-7317 ISSN 0027-8424.

*Communication,* 114, 2, (July 1983), pp. 737-742, ISSN 0006-291X.

2002), pp. G1352-1359, ISSN 0193-1857.

2001), pp. 14134-14142, ISSN 0006-2960.

2008), pp. 5821-5832, ISSN 0950-9232.

(August 2006), pp. 13086-13091, ISNN 0027-8424.

ISSN 0021-9533.

vascular smooth muscle cells through telomerase activation*. Molecular and Cellular* 

Pueyo, C. (1996). The levels of ribonucleotide reductase, thioredoxin, glutaredoxin 1, and GSH are balanced in Escherichia coli K12. *Journal of Biology Chemistry*, 271,

thioredoxin redox during differentiation in human colon epithelial (Caco-2) cells. *American journal of physiology: Gastrointestinal and liver physiology*, 283, 6, (December

tumorigenesis*. Journal of Cell Science*, 113, Pt 22, (November 2000), pp. 3897-905,

transformed cells: the possible role of superoxide and hydrogen peroxide. *Medical* 

identification, and significance of DNA-protein cross-links in mammalian cells. *The British Journal of Cancer Supplement,* 8, (June 1987), pp. 135-140, ISSN 0306-9443. Painter, R. B. & Young, B. R. (1980). Radiosensitivity in ataxia-telangiectasia: A new

explanation. *Proceedings of the National Academy of Sciences of the United States of* 

signaling mechanism for density-dependent inhibition of cell growth. *Journal of Biology Chemistry*, 275, 49, (December 2000), pp. 38891-38899, ISSN 0042-6989. Pineda-Molina, E.; Klatt, P.; Vázquez, J.; Marina, A.; García de Lacoba, M.; Pérez-Sala, D.

Lamas, S. (2001). Glutathionylation of the p50 subunit of NF-kappaB: a mechanism for redox-induced inhibition of DNA binding. *Biochemistry*, 40, 47, (November

glutathione content in A549 cells. *Biochemical and Biophysical Research* 

Reddy, S.P. (2008). Genetic disruption of the Nrf2 compromises cell-cycle progression by impairing GSH-induced redox signaling. *Oncogene*, 27, 44, (October

Heintz, N.H.; Heim, J.; Ho, Y.S.; Matthews, D.E.; Wouters, E.F. Janssen-Heininger, Y.M. (2006). Dynamic redox control of NF-kappaB through glutaredoxin-regulated S-glutathionylation of inhibitory kappaB kinase beta. *Proceedings of the National Academy of Sciences of the United States of America*, 103, 35,


**13** 

*Japan* 

**Role for PKCδ on Apoptosis in the** 

*Medical Research Institute, Tokyo Medical and Dental University, Tokyo,* 

Genotoxic stress induces cell cycle arrest, DNA repair, and apoptotic cell death. The decision by cells either to repair DNA lesions and continue through the cell cycle or to undergo apoptosis is relevant to the incidence of mutagenesis and, subsequently, carcinogenesis. In this regard, incomplete repair of DNA damage prior to replication or mitosis can result in the accumulation of heritable genetic changes. Therapeutic anti-cancer treatments that use genotoxic agents must strike a balance between induction of repair and apoptosis in order to maximize the therapeutic effect. However, the nature of the cellular signaling response that determines cell fate such as survival or death is far from being understood. Certain insights have been derived from the finding that diverse isozymes of the protein kinase C (PKC) family are activated in response to DNA damage. PKC-mediated signaling pathway modulates destiny of cells following genotoxic insults (Yoshida 2007a, Yoshida 2008a). In particular, recent studies have shown that certain isozyme of PKC controls function of the p53 tumor suppressor in induction of cell cycle arrest, DNA repair, and apoptosis. In the past 10 years, understanding the molecular mechanisms of apoptosis mediated by PKC has advanced considerably, and the primary focus of this review is to provide an overview of PKC and p53,

its mode of action and its physiological role in DNA damage-induced apoptosis.

The protein kinase C (PKC) family of serine-threonine kinases was first described as a calcium-activated, phospholipid-dependent serine/threonine protein kinase (Takai et al. 1977). PKC is activated diacylglycerol (DAG) hydrolyzed from phosphatidylinositol (PI) by phospholipase C (PLC) under a different cell-signaling system (Nishizuka 1984, Nishizuka 1988, Nishizuka 1992, Nishizuka 1995). It has attracted attention as an intracellular receptor for tumor-promotor phorbol esters, such as 12-O-tetradecanoyl-13-phorbol acetate (TPA) (Niedel et al. 1983). Although PKC had been recognized as a protein kinase, subsequent studies have revealed that it belongs to a family of serine/threonine-specific protein kinases and is activated by diverse stimuli and participates in various cellular processes, such as growth, differentiation, apoptosis, and cellular senescence (Casabona 1997, Clemens et al. 1992, Goodnight et al. 1994, Hofmann 1997, Hug and Sarre 1993, Nishizuka 1984, Nishizuka 1988, Nishizuka 1992, Nishizuka 1995). PKC consists of at least 11 isozymes (, I, II, , , , , , , / and ) with selective tissue distribution, activators, and substrates. PKC isozymes

**1. Introduction** 

**2. Protein kinase C** 

**DNA Damage Response** 

Kiyotsugu Yoshida

