Induction of apoptosis by cytoplasmically localized wild-type p53 and the S121F mutant super p53

Yasuda,Katsuhiro; Kato,Shunsuke; Sakamoto,Yasuhiro; Watanabe,Gou; Mashiko,Satsuki; Sato,Atsuko; Kakudo,Yuichi; Ishioka,Chikashi

doi:10.3892/ol.2012.624

Induction of apoptosis by cytoplasmically localized wild-type p53 and the S121F mutant super p53

Authors:
- Katsuhiro Yasuda
- Shunsuke Kato
- Yasuhiro Sakamoto
- Gou Watanabe
- Satsuki Mashiko
- Atsuko Sato
- Yuichi Kakudo
- Chikashi Ishioka
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Affiliations: Department of Clinical Oncology, Institute of Development, Aging and Cancer, Tohoku University, Sendai, Miyagi 980-8575, Japan
Published online on: February 29, 2012 https://doi.org/10.3892/ol.2012.624
Pages: 978-982

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Abstract

After DNA damage, p53 is accumulated in the nucleus and transactivates downstream genes and induces apoptosis. There are two pathways in p53-dependent apoptosis, the transactivation-dependent and -independent pathway. In this study, we constructed p53-inducible glioblastoma cell lines and analyzed them for the induction of apoptosis and transactivation of p53-downstream genes after the nuclear or cytoplasmic expression of p53. To sequester p53 in the cytoplasm, we used p53 mutant with arginine to glycine substitution at residue 306 (R306G). Wild-type p53 retained the ability to arrest the cell cycle, and a p53 mutant with serine to phenylalanine substitution at residue 121 (S121F), which has a strong ability to induce apoptosis, retained this ability even when both the wild-type and p53 and S121F mutant were exclusively sequestered from the nucleus into the cytoplasm. Notably, cytoplasmically sequestered wild-type p53 and S121F mutant transactivated the downstream genes with distinct expression profiles, and the strong apoptotic ability of S121F was not associated with its transactivation activity. These results underscore the existence of transactivation-independent apoptosis and cytoplasmic function of p53.

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1	Hollstein M, Rice K, Greenblatt MS, et al: Database of p53 gene somatic mutations in human tumors and cell lines. Nucleic Acids Res. 22:3551–3555. 1994.PubMed/NCBI
2	Soussi T: p53 alterations in human cancer: more questions than answers. Oncogene. 26:2145–2156. 2007. View Article : Google Scholar : PubMed/NCBI
3	Harris SL and Levine AJ: The p53 pathway: positive and negative feedback loops. Oncogene. 24:2899–2908. 2005. View Article : Google Scholar : PubMed/NCBI
4	Soussi T and Lozano G: p53 mutation heterogeneity in cancer. Biochem Biophys Res Commun. 331:834–842. 2005. View Article : Google Scholar : PubMed/NCBI
5	Moll UM, LaQuaglia M, Benard J and Riou G: Wild-type p53 protein undergoes cytoplasmic sequestration in undifferentiated neuroblastomas but not in differentiated tumors. Proc Natl Acad Sci USA. 92:4407–4411. 1995. View Article : Google Scholar : PubMed/NCBI
6	Moll UM, Riou G and Levine AJ: Two distinct mechanisms alter p53 in breast cancer: mutation and nuclear exclusion. Proc Natl Acad Sci USA. 89:7262–7266. 1992. View Article : Google Scholar : PubMed/NCBI
7	Nagpal J, Jamoona A, Gulati ND, et al: Revisiting the role of p53 in primary and secondary glioblastomas. Anticancer Res. 26:4633–4639. 2006.PubMed/NCBI
8	Liang SH and Clarke MF: A bipartite nuclear localization signal is required for p53 nuclear import regulated by a carboxyl-terminal domain. J Biol Chem. 274:32699–32703. 1999. View Article : Google Scholar : PubMed/NCBI
9	Kim IS, Kim DH, Han SM, et al: Truncated form of importin alpha identified in breast cancer cell inhibits nuclear import of p53. J Biol Chem. 275:23139–23145. 2000. View Article : Google Scholar : PubMed/NCBI
10	Rodriguez-Lopez AM, Xenaki D, Eden TO, Hickman JA and Chresta CM: MDM2 mediated nuclear exclusion of p53 attenuates etoposide-induced apoptosis in neuroblastoma cells. Mol Pharmacol. 59:135–143. 2001.PubMed/NCBI
11	Kaul SC, Deocaris CC and Wadhwa R: Three faces of mortalin: a housekeeper, guardian and killer. Exp Gerontol. 42:263–274. 2007. View Article : Google Scholar : PubMed/NCBI
12	Andrews P, He YJ and Xiong Y: Cytoplasmic localized ubiquitin ligase cullin 7 binds to p53 and promotes cell growth by antagonizing p53 function. Oncogene. 25:4534–4548. 2006. View Article : Google Scholar : PubMed/NCBI
13	Nikolaev AY, Li M, Puskas N, Qin J and Gu W: Parc: a cytoplasmic anchor for p53. Cell. 112:29–40. 2003. View Article : Google Scholar : PubMed/NCBI
14	Kato S, Han SY, Liu W, et al: Understanding the function-structure and function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis. Proc Natl Acad Sci USA. 100:8424–8429. 2003. View Article : Google Scholar
15	Freeman J, Schmidt S, Scharer E and Iggo R: Mutation of conserved domain II alters the sequence specificity of DNA binding by the p53 protein. EMBO J. 13:5393–5400. 1994.PubMed/NCBI
16	Saller E, Tom E, Brunori M, et al: Increased apoptosis induction by 121F mutant p53. EMBO J. 18:4424–4437. 1999. View Article : Google Scholar : PubMed/NCBI
17	Kakudo Y, Shibata H, Otsuka K, Kato S and Ishioka C: Lack of correlation between p53-dependent transcriptional activity and the ability to induce apoptosis among 179 mutant p53s. Cancer Res. 65:2108–2114. 2005. View Article : Google Scholar : PubMed/NCBI
18	Goldman SC, Chen CY, Lansing TJ, Gilmer TM and Kastan MB: The p53 signal transduction pathway is intact in human neuroblastoma despite cytoplasmic localization. Am J Pathol. 148:1381–1385. 1996.
19	Chen X, Ko LJ, Jayaraman L and Prives C: p53 levels, functional domains, and DNA damage determine the extent of the apoptotic response of tumor cells. Genes Dev. 10:2438–2451. 1996. View Article : Google Scholar : PubMed/NCBI
20	Haupt Y, Rowan S, Shaulian E, Vousden KH and Oren M: Induction of apoptosis in HeLa cells by trans-activation-deficient p53. Genes Dev. 9:2170–2183. 1995. View Article : Google Scholar : PubMed/NCBI