p21CDKN1A participates in base excision repair by regulating the activity of poly(ADP-ribose) polymerase-1 |
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| Authors: | Ornella Cazzalini Francesca Donà Monica Savio Micol Tillhon Cristina Maccario Paola Perucca Lucia A Stivala A Ivana Scovassi Ennio Prosperi |
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| Institution: | 1. Dipartimento di Medicina Sperimentale, Università di Pavia, Pavia, Italy;2. Istituto di Genetica Molecolare del CNR (IGM-CNR), Pavia, Italy;1. Max Planck Institute for Biology of Ageing, Joseph-Stelzmann-Strasse 9b, Cologne 50931, Germany;2. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK;1. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK;1. Nihon Superior Centre for the Manufacture of Electronic Materials (NS CMEM), School of Mechanical and Mining Engineering, The University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia;2. Department of Materials, Imperial College London, London SW7 2AZ, UK;3. Australian Synchrotron, ANSTO, Clayton, VIC 3168, Australia;4. Department of Materials Science and Engineering, Kyoto University, Sakyo, Kyoto 606-8501, Japan;5. Department of Applied Quantum Physics and Nuclear Engineering, Kyushu University, Fukuoka 819-0395, Japan;3. From the School of Biomedical Sciences, LKS Faculty of Medicine, L1–46, Laboratory Block, 21 Sassoon Road, Pokfulam, Hong Kong;5. the State Key Laboratory of Brain and Cognitive Sciences, University of Hong Kong, L1–46, Laboratory Block, 21 Sassoon Road, Pokfulam, Hong Kong and;4. the Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213 |
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| Abstract: | The cell cycle inhibitor p21CDKN1A has been shown to participate in nucleotide excision repair by interacting with PCNA. Here we have investigated whether p21 plays a role in base excision repair (BER), by analyzing p21 interactions with BER factors, and by assessing the response of p21?/? human fibroblasts to DNA damage induced by alkylating agents. Absence of p21 protein resulted in a higher sensitivity to alkylation-induced DNA damage, as indicated by reduced clonogenic efficiency, defective DNA repair (assessed by the comet test), and by persistence of histone H2AX phosphorylation. To elucidate the mechanisms at the basis of the function of p21 in BER, we focused on its interaction with poly(ADP-ribose) polymerase-1 (PARP-1), an important player in this repair process. p21 was found to bind the automodification/DNA binding domain of PARP-1, although some interaction occurred also with the catalytic domain after DNA damage. This association was necessary to regulate PARP-1 activity since poly(ADP-ribosylation) induced by DNA damage was higher in p21?/? human fibroblasts than in parental p21+/+ cells, and in primary fibroblasts after p21 knock-down by RNA interference. Concomitantly, recruitment of PARP-1 and PCNA to damaged DNA was greater in p21?/? than in p21+/+ fibroblasts. This accumulation resulted in persistent interaction of PARP-1 with BER factors, such as XRCC1 and DNA polymerase β, suggesting that prolonged association reduced the DNA repair efficiency. These results indicate that p21 regulates the interaction between PARP-1 and BER factors, to promote efficient DNA repair. |
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