**8. References**

298 Selected Topics in DNA Repair

also in amplifying upstream caspase signaling. Most of the studies mentioned above suggest that the role of PKC in the induction of apoptosis is tightly associated with its caspasedependent cleavage and the regulation of p53. However, functional regulation of p53 by PKC remains largely unclear. In this regard, thorough investigation coupled with PKC and p53 should be enhanced from multiple views. In the encounter with genotoxic insults, ATM controls various cellular responses, such as cell cycle arrest, transcription, DNA repair, and apoptosis. In this context, DNA damage-induced PKC is modulated under ATM, suggesting the notion that establishment of the ATMPKCp53 signaling cascade provides new mechanistic light on how PKC functions as the pro-apoptotic kinase in the nucleus (Figure 1) (Yoshida 2007a, Yoshida 2008a). While dysregulation of the PKC signalsome confers resistance to anticancer drugs (Meinhardt et al. 1999), there is little

Fig. 1. A hypothetical schema for nuclear targeting of PKC in response to DNA damage. Following DNA damage, PKC translocates from the cytoplasm into the nucleus. In

addition, some genotoxic stress also exerts cytoplasmic oxidative stress to activate PKC. In the nucleus, PKC is activated by ATM, then induces apoptosis (or DNA repair) in a p53-

dependent manner.


Role for PKCδ on Apoptosis in the DNA Damage Response 301

Humphries, M. J., Limesand, K. H., Schneider, J. C., Nakayama, K. I., Anderson, S. M.,

Joseloff, E., Cataisson, C., Aamodt, H. et al. (2002). Src family kinases phosphorylate protein

Kaul, S., Anantharam, V., Yang, Y., Choi, C. J., Kanthasamy, A., Kanthasamy, A. G. (2005).

Konishi, H., Yamauchi, E., Taniguchi, H. et al. (2001). Phosphorylation sites of protein kinase

Kubbutat, M. H., Vousden, K. H. (1998). Keeping an old friend under control: regulation of

Leitges, M., Mayr, M., Braun, U. et al. (2001). Exacerbated vein graft arteriosclerosis in

Liu, H., Lu, Z. G., Miki, Y., Yoshida, K. (2007). Protein kinase C delta induces transcription

Lu, Z., Hornia, A., Jiang, Y. W., Zang, Q., Ohno, S., Foster, D. A. (1997). Tumor promotion by

Magnelli, L., Cinelli, M., Chiarugi, V. (1995). Phorbol esters attenuate the expression of p53

Majumder, P. K., Pandey, P., Sun, X. et al. (2000). Mitochondrial translocation of protein

Matassa, A. A., Carpenter, L., Biden, T. J., Humphries, M. J., Reyland, M. E. (2001). PKCdelta

Matsuda, K., Yoshida, K., Taya, Y., Nakamura, K., Nakamura, Y., Arakawa, H. (2002). p53AIP1 regulates the mitochondrial apoptotic pathway. Cancer Res 62:2883-9. Meek, D. W. (1998). Multisite phosphorylation and the integration of stress signals at p53.

Meinhardt, G., Roth, J., Totok, G., Auner, H., Emmerich, B., Hass, R. (1999). Signaling defect

Niedel, J. E., Kuhn, L. J., Vandenbark, G. R. (1983). Phorbol diester receptor copurifies with

Nigro, J. M., Baker, S. J., Preisinger, A. C. et al. (1989). Mutations in the p53 gene occur in

Nishizuka, Y. (1984). The role of protein kinase C in cell surface signal transduction and

associated with resistance to apoptosis. Exp Cell Res 247:534-42.

protein kinase C. Proc Natl Acad Sci U S A 80:36-40.

diverse human tumour types. Nature 342:705-8.

tumour promotion. Nature 308:693-8.

Cdelta in dopaminergic neuronal cells. J Biol Chem 280:28721-30.

protein kinase Cdelta-null mice. J Clin Invest 108:1505-12.

apoptotic response to DNA damage. Mol Cell Biol 27:8480-91.

depleting cells of protein kinase C delta. Mol Cell Biol 17:3418-28.

mouse in vivo. J Biol Chem 281:9728-37.

keratinocytes. J Biol Chem 277:12318-23.

Natl Acad Sci U S A 98:6587-92.

p53 stability. Mol Med Today 4:250-6.

Biochem Biophys Res Commun 215:641-5.

Chem 275:21793-6.

Chem 276:29719-28.

Cell Signal 10:159-66.

Reyland, M. E. (2006). Suppression of apoptosis in the protein kinase Cdelta null

kinase C delta on tyrosine residues and modify the neoplastic phenotype of skin

Tyrosine phosphorylation regulates the proteolytic activation of protein kinase

C delta in H2O2-treated cells and its activation by tyrosine kinase in vitro. Proc

of the TP53 tumor suppressor gene by controlling death-promoting factor Btf in the

in cells treated with doxorubicin and protect TS-P53/K562 from apoptosis.

kinase C delta in phorbol ester-induced cytochrome c release and apoptosis. J Biol

is required for mitochondrial-dependent apoptosis in salivary epithelial cells. J Biol

in the activation of caspase-3 and PKCdelta in human TUR leukemia cells is


Denning, M. F., Dlugosz, A. A., Threadgill, D. W., Magnuson, T., Yuspa, S. H. (1996). Activation

tyrosine phosphorylation of protein kinase C delta. J Biol Chem 271:5325-31. DeVries, T. A., Neville, M. C., Reyland, M. E. (2002). Nuclear import of PKCdelta is

DeVries-Seimon, T. A., Ohm, A. M., Humphries, M. J., Reyland, M. E. (2007). Induction of

Donehower, L. A., Harvey, M., Slagle, B. L. et al. (1992). Mice deficient for p53 are

Eitel, K., Staiger, H., Rieger, J. et al. (2003). Protein kinase C delta activation and

Emoto, Y., Manome, Y., Meinhardt, G. et al. (1995). Proteolytic activation of protein kinase C

Endo, K., Oki, E., Biedermann, V. et al. (2000). Proteolytic cleavage and activation of

Fournier, A., Murray, A. W. (1987). Application of phorbol ester to mouse skin causes a

Ghayur, T., Hugunin, M., Talanian, R. V. et al. (1996). Proteolytic activation of protein kinase

Ghosh, J. C., Suzuki, K., Kodama, S., Watanabe, M. (1999). Effects of protein kinase

Goodnight, J., Mischak, H., Mushinski, J. F. (1994). Selective involvement of protein kinase C

Hansen, L. A., Monteiro-Riviere, N. A., Smart, R. C. (1990). Differential down-regulation of

Hew, H. C., Liu, H., Miki, Y., Yoshida, K. (2011). PKCdelta regulates Mdm2 independently

Hofmann, J. (1997). The potential for isoenzyme-selective modulation of protein kinase C.

Hug, H., Sarre, T. F. (1993). Protein kinase C isoenzymes: divergence in signal transduction?

of p53 in the apoptotic response to DNA damage. Mol Carcinog.

delta by an ICE-like protease in apoptotic cells. EMBO J 14:6148-56.

rapid and sustained loss of protein kinase C. Nature 330:767-9.

cells following X-irradiation. J Radiat Res (Tokyo) 40:23-37.

21:6050-60.

282:22307-14.

275:18476-81.

Med 184:2399-404.

Cancer Res 50:5740-5.

FASEB J 11:649-69.

Biochem J 291 ( Pt 2):329-43.

209.

secreting cells. Diabetes 52:991-7.

21.

of the epidermal growth factor receptor signal transduction pathway stimulates

required for apoptosis: identification of a novel nuclear import sequence. EMBO J

apoptosis is driven by nuclear retention of protein kinase C delta. J Biol Chem

developmentally normal but susceptible to spontaneous tumours. Nature 356:215-

translocation to the nucleus are required for fatty acid-induced apoptosis of insulin-

protein kinase C [micro] by caspase-3 in the apoptotic response of cells to 1 beta -D-arabinofuranosylcytosine and other genotoxic agents. J Biol Chem

C delta by an ICE/CED 3-like protease induces characteristics of apoptosis. J Exp

inhibitors on the accumulation kinetics of p53 protein in normal human embryo

isozymes in differentiation and neoplastic transformation. Adv Cancer Res 64:159-

epidermal protein kinase C by 12-O-tetradecanoylphorbol-13-acetate and diacylglycerol: association with epidermal hyperplasia and tumor promotion.


Role for PKCδ on Apoptosis in the DNA Damage Response 303

Vogelstein, B., Lane, D., Levine, A. J. (2000). Surfing the p53 network. Nature 408:307-10. Wang, Q. J., Bhattacharyya, D., Garfield, S., Nacro, K., Marquez, V. E., Blumberg, P. M.

Watanabe, T., Ono, Y., Taniyama, Y. et al. (1992). Cell division arrest induced by phorbol

Yamaguchi, T., Kimura, J., Miki, Y., Yoshida, K. (2007a). The deubiquitinating enzyme

Yamaguchi, T., Miki, Y., Yoshida, K. (2007b). Protein kinase C delta activates IkappaB-kinase

Yoshida, K. (2007a). PKCdelta signaling: Mechanisms of DNA damage response and

Yoshida, K. (2007b). Regulation for nuclear targeting of the Abl tyrosine kinase in response

Yoshida, K. (2008a). Nuclear trafficking of pro-apoptotic kinases in response to DNA

Yoshida, K. (2008b). Role for DYRK family kinases on regulation of apoptosis. Biochem

Yoshida, K., Kufe, D. (2001). Negative regulation of the SHPTP1 protein tyrosine

Yoshida, K., Liu, H., Miki, Y. (2006a). Protein Kinase C delta regulates Ser46

Yoshida, K., Miki, Y. (2005). Enabling death by the Abl tyrosine kinase: mechanisms for nuclear shuttling of c-Abl in response to DNA damage. Cell Cycle 4:777-9. Yoshida, K., Miki, Y., Kufe, D. (2002). Activation of SAPK/JNK signaling by protein kinase

Yoshida, K., Wang, H. G., Miki, Y., Kufe, D. (2003). Protein kinase Cdelta is responsible for

Yoshida, K., Yamaguchi, T., Natsume, T., Kufe, D., Miki, Y. (2005). JNK phosphorylation of

Yoshida, K., Yamaguchi, T., Shinagawa, H., Taira, N., Nakayama, K. I., Miki, Y. (2006b).

Cdelta in response to DNA damage. J Biol Chem 277:48372-8.

death in response to DNA damage. Mol Cell Biol 26:3414-31.

phosphatase by protein kinase C delta in response to DNA damage. Mol Pharmacol

phosphorylation of p53 tumor suppressor in the apoptotic response to DNA

constitutive and DNA damage-induced phosphorylation of Rad9. EMBO J 22:1431-

14-3-3 proteins regulates nuclear targeting of c-Abl in the apoptotic response to

Protein kinase C delta activates topoisomerase IIalpha to induce apoptotic cell

Biol Chem 252:7603-9.

Chem 274:37233-9.

Acad Sci U S A 89:10159-63.

apoptosis. Cell Signal 19:892-901.

damage. Trends Mol Med 14:305-13.

damage. J Biol Chem 281:5734-40.

DNA damage. Nat Cell Biol 7:278-85.

Pharmacol 76:1389-1394.

60:1431-8.

41.

to DNA damage. Adv Exp Med Biol 604:155-65.

Biol Chem 282:33943-8.

Signal 19:2088-97.

Purification and characterization of an active enzyme from bovine cerebellum. J

(1999). Differential localization of protein kinase C delta by phorbol esters and related compounds using a fusion protein with green fluorescent protein. J Biol

ester in CHO cells overexpressing protein kinase C-delta subspecies. Proc Natl

USP11 controls an IKKalpha -p53 signaling pathway in response to TNFalpha. J

alpha to induce the p53 tumor suppressor in response to oxidative stress. Cell


Nishizuka, Y. (1988). The molecular heterogeneity of protein kinase C and its implications

Nishizuka, Y. (1992). Intracellular signaling by hydrolysis of phospholipids and activation of

Nishizuka, Y. (1995). Protein kinase C and lipid signaling for sustained cellular responses.

Oda, K., Arakawa, H., Tanaka, T. et al. (2000). p53AIP1, a potential mediator of p53-

Okamura, S., Arakawa, H., Tanaka, T. et al. (2001). p53DINP1, a p53-inducible gene,

Oren, M. (1999). Regulation of the p53 tumor suppressor protein. J Biol Chem 274:36031-

Oren, M., Damalas, A., Gottlieb, T. et al. (2002). Regulation of p53: intricate loops and

Raman, V., Martensen, S. A., Reisman, D. et al. (2000). Compromised HOXA5 function can

Reddig, P. J., Dreckschmidt, N. E., Ahrens, H. et al. (1999). Transgenic mice overexpressing

Ryan, K. M., Phillips, A. C., Vousden, K. H. (2001). Regulation and function of the p53 tumor

Scheel-Toellner, D., Pilling, D., Akbar, A. N. et al. (1999). Inhibition of T cell apoptosis by

Shieh, S. Y., Ikeda, M., Taya, Y., Prives, C. (1997). DNA damage-induced phosphorylation of

Siliciano, J. D., Canman, C. E., Taya, Y., Sakaguchi, K., Appella, E., Kastan, M. B. (1997).

Stuart, E. T., Haffner, R., Oren, M., Gruss, P. (1995). Loss of p53 function through PAX-

Taira, N., Nihira, K., Yamaguchi, T., Miki, Y., Yoshida, K. (2007). DYRK2 is targeted to the

Taira, N., Yamamoto, H., Yamaguchi, T., Miki, Y., Yoshida, K. (2010). ATM augments

Takai, Y., Kishimoto, A., Inoue, M., Nishizuka, Y. (1977). Studies on a cyclic nucleotide-

Reyland, M. E. (2007). Protein kinase Cdelta and apoptosis. Biochem Soc Trans 35:1001-4. Reyland, M. E., Anderson, S. M., Matassa, A. A., Barzen, K. A., Quissell, D. O. (1999). Protein

protein kinase Cdelta in the epidermis are resistant to skin tumor promotion by 12-

kinase C delta is essential for etoposide-induced apoptosis in salivary gland acinar

IFN-beta rapidly reverses nuclear translocation of protein kinase C-delta. Eur J

DNA damage induces phosphorylation of the amino terminus of p53. Genes Dev

nucleus and controls p53 via Ser46 phosphorylation in the apoptotic response to

nuclear stabilization of DYRK2 by inhibiting MDM2 in the apoptotic response to

independent protein kinase and its proenzyme in mammalian tissues. I.

limit p53 expression in human breast tumours. Nature 405:974-8.

O-tetradecanoylphorbol-13-acetate. Cancer Res 59:5710-8.

regulates p53-dependent apoptosis. Mol Cell 8:85-94.

delicate balances. Biochem Pharmacol 64:865-71.

suppressor protein. Curr Opin Cell Biol 13:332-7.

p53 alleviates inhibition by MDM2. Cell 91:325-34.

mediated transcriptional repression. EMBO J 14:5638-45.

cells. J Biol Chem 274:19115-23.

DNA damage. Mol Cell 25:725-38.

DNA damage. J Biol Chem 285:4909-19.

Immunol 29:2603-12.

11:3471-81.

dependent apoptosis, and its regulation by Ser-46-phosphorylated p53. Cell

for cellular regulation. Nature 334:661-5.

protein kinase C. Science 258:607-14.

FASEB J 9:484-96.

102:849-62.

4.

Purification and characterization of an active enzyme from bovine cerebellum. J Biol Chem 252:7603-9.


**14** 

*Russia* 

**New Players in Recognition** 

Svetlana Khodyreva and Olga Lavrik

**of Intact and Cleaved AP Sites:** 

*Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences,* 

**Implication in DNA Repair in Mammalian Cells** 

The apurinic/apyrimidinic (AP) sites (called also abasic sites) are common lesion in genomic DNA, arising at a frequency of 10,000 to 50,000 lesions per mammalian cell per day (Lindahl, 1993). Unrepaired AP sites present mutagenic and cytotoxic consequences to the cell (Wilson; & Thompson, 1997). Most of the abasic sites are believed to result directly from spontaneous depurination, or indirectly from deamination of cytosine to uracil, which is then eliminated by uracil glycosylases. AP sites also result from hydrolysis of oxidized or alkylated bases by lesion-specific glycosylases during the early stage of base excision repair (BER) (McCullough et al., 1999). AP sites in isolated DNA are rather stable, but can be converted to single-strand breaks by alkali treatment, heating or nucleophilic attack at the aldehydic C1' group (Burrows & Muller, 1998). Intact abasic sites are noncoding lesions and *in vivo* can be stable enough to be mutagenic during DNA replication (Loeb & Preston, 1986). To protect genome integrity, eukaryotic organisms have robust enzyme activities, mainly APE1 in mammalian cells (Wilson & Barsky, 2001), that recognize abasic sites and cut the DNA backbone initiating the repair process. The continuous generation and repair of AP sites results in a steady-state levels of AP sites in mammalian cells in the range of approximately 1 site per 106 nucleotides (Atamna et al., 2000, Mohsin Ali et al., 2004). The number of AP sites can increase dramatically under stressful conditions such as X-ray or UV light irradiation or oxidative and alkylating agent exposure (Atamna et al., 2000). Considering the ubiquity of these lesions, it is reasonable to assume that wide range of cellular proteins can interact with abasic sites depending on the

The chapter is devoted to search of previously unrecognized proteins capable to interact with intact or cleaved AP sites. We mainly focused on proteins that form Schiff base upon this interaction. In most cases, these proteins are able to process AP sites although less efficiently than previously known counterparts. The biological role of these interactions in

In genomic DNA of higher eukaryotes AP sites irrespectively of their origin are thought to

providing of backup pathways of DNA repair processes is also discussed.

be repaired by base excision repair (Almeida & Sobol, 2007; Hegde et al., 2008).

**1. Introduction** 

physiological state and stages of cell cycle.

**2. Proteins that recognize AP sites** 

Yuan, Z. M., Utsugisawa, T., Ishiko, T. et al. (1998). Activation of protein kinase C delta by the c-Abl tyrosine kinase in response to ionizing radiation. Oncogene 16:1643-8.
