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  • Original Paper
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Functional analysis and intracellular localization of p53 modified by SUMO-1

Oncogene volume 20, pages 2587–2599 (2001)Cite this article

Abstract

p53 tumor suppressor is a subject of several post-translational modifications, including phosphorylation, ubiquitination and acetylation, which regulate p53 function. A new covalent modification of p53 at lysine 386 by SUMO-1 was recently identified. To elucidate the function of sumoylated p53, we compared the properties of wild type p53 and sumoylation-deficient p53 mutant, K386R. No differences were found between wild type p53 and K386R mutant of p53 in transactivation or growth suppression assays. Moreover, overexpression of SUMO-1 has no effect on p53-regulated transcription. Biochemical fractionation showed that sumoylated p53 is localized in the nucleus and is tightly bound to chromatin structures. p53 and SUMO-1 co-localized in PML nuclear bodies in 293 cells and the nucleoli in MCF7 and HT1080 cells. However, sumoylation-deficient p53 mutant showed a similar pattern of intranuclear localization, suggesting that SUMO-1 does not target p53 to subnuclear structures. These data indicate that SUMO-1 modification of p53 at lysine 386 may not be essential for p53's cellular localization, transcriptional activation, or growth regulation.

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Abbreviations

SUMO-1:

Small Ubiquitin Modifier 1

PML:

Promyelocytic Leukemia Protein

CAT:

chloramphenicol acetyl transferase

HA:

hemagglutinin

NEM:

N-ethylmaleimide

DMEM:

Dulbecco's Modified Eagle's Medium

PBS:

phosphate-buffered saline

DAPI:

4,6-diamidino-2-phenylindole

GFP:

green fluorescence protein.

References

  • Ashcroft M, Kubbutat MH, Vousden KH . 1999 Mol. Cell. Biol. 19: 1751–1758

  • Avantaggiati ML, Ogryzko V, Gardner K, Giordano A, Levine AS, Kelly K . 1997 Cell 89: 1175–1184

  • Bates S, Phillips AC, Clark PA, Stott F, Peters G, Ludwig RL, Vousden KH . 1998 Nature 395: 124–125

  • Blattner C, Tobiasch E, Litfen M, Rahmsdorf HJ, Herrlich P . 1999 Oncogene 18: 1723–1732

  • Boddy MN, Howe K, Etkin LD, Solomon E, Freemont PS . 1996 Oncogene 13: 971–982

  • Buschmann T, Fuchs SY, Lee CG, Pan ZQ, Ronai Z . 2000 Cell 101: 753–762

  • Carter T, James C, Chan E, Greene B . 1987 Cancer Res. 47: 803–808

  • Chehab NH, Malikzay A, Appel M, Halazonetis TD . 2000 Genes Dev. 14: 278–288

  • Debbas M, White E . 1993 Genes Dev. 7: 546–554

  • Desterro JM, Thomson J, Hay RT . 1997 FEBS Lett. 417: 297–300

  • Desterro JM, Rodriguez MS, Hay RT . 1998 Mol. Cell 2: 233–239

  • Finley Jr RL, Brent R . 1995 Interaction Trap Cloning with yeast In: DNA Cloning, Expression Systems: A Practical Approach, Hames BD and Glover DM (ed.) Oxford Univ. Press, Oxford: pp. 169–203

  • Fiscella M, Ullrich SJ, Zambrano N, Shields MT, Lin D, Lees-Miller SP, Anderson CW, Mercer WE, Appella E . 1993 Oncogene 8: 1519–1528

  • Fuchs B, O'Connor D, Fallis L, Scheidtmann KH, Lu X . 1999 Oncogene 10: 789–793

  • Gerace L, Ottaviano Y, Kondor-Koch C . 1982 J. Cell Biol. 95: 826–837

  • Giaccia AJ, Kastan MB . 1998 Genes Dev. 12: 2973–2983

  • Gong L, Millas S, Maul GG, Yeh ET . 2000 J. Biol. Chem. 275: 3355–3359

  • Gostissa M, Hengstermann A, Fogal V, Sandy P, Schwarz SE, Scheffner M, Del Sal G . 1999 EMBO J. 18: 6462–6471

  • Graham JM, Rickwood D . 1997 Subcellular Fractionation: A Practical Approach OIRL Press, Oxford University Press Inc.: NY p. 71–84

  • Graham FL, Smiley J, Russell WC, Nairn R . 1977 J. Gen. Virol. 36: 59–74

  • Gu W, Shi XL, Roeder RG . 1997 Nature 387: 819–823

  • Haupt Y, Maya R, Kazaz A, Oren M . 1997 Nature 387: 296–299

  • Hollstein M, Shomer B, Greenblatt M, Soussi T, Hovig E, Montesano R, Harris CC . 1996 Nucleic Acids Res. 24: 141–146

  • Holtz D, Tanaka RA, Hartwig J, McKeon F . 1989 Cell 59: 969–977

  • Honda R, Tanaka H, Yasuda H . 1997 FEBS Lett. 420: 25–27

  • Johnson ES, Blobel G . 1997 J. Biol. Chem. 272: 26799–26802

  • Johnson ES, Schwienhorst I, Dohmen RJ, Blobel G . 1997 EMBO J. 16: 5509–5519

  • Kamitani T, Kito K, Nguyen HP, Wada H, Fukuda-Kamitani T, Yeh ET . 1998 J. Biol. Chem. 273: 26675–26682

  • Kamitani T, Nguyen HP, Yeh ET . 1997 J. Biol. Chem. 272: 14001–14004

  • Kay RR, Fraser D, Johnston IR . 1972 Eur. J. Biochem. 30: 145–154

  • Kim YH, Choi CY, Kim Y . 1999 Proc. Natl. Acad. Sci. USA 96: 12350–12355

  • Kirsch DG, Kastan MB . 1998 J. Clin. Oncol. 16: 3158–3168

  • Kondratov RV, Kuznetsov NV, Pugacheva EN, Almazov VP, Prasolov VS, Kopnin BP, Chumakov PM . 1996 Mol. Biol. (Mosk) 30: 613–620

  • Kubbutat MH, Vousden KH . 1998 Mol. Med. Today 4: 250–256

  • Lain S, Midgley C, Sparks A, Lane EB, Lane DP . 1999 Exp. Cell Res. 248: 457–472

  • Lakin ND, Jackson SP . 1999 Oncogene 18: 7644–7655

  • Leppard KN, Everett RD . 1999 J. Gen. Virol. 80: 997–1008

  • Li SJ, Hochstrasser M . 1999 Nature 398: 246–251

  • Li W, Hesabi B, Babbo A, Pacione C, Liu J, Chen DJ, Nickoloff JA, Shen Z . 2000 Nuclei Acids Res. 28: 1145–1153

  • Lill NL, Grossman SR, Ginsberg D, DeCaprio J, Livingston DM . 1997 Nature 387: 823–827

  • Lindström MS, Klangby U, Inoue R, Pisa P, Wiman KG, Asker CE . 2000 Exp. Cell Res. 256: 400–410

  • Liu L, Scolnick DM, Trievel RC, Zhang HB, Marmorstein R, Halazonetis TD, Berger SL . 1999 Mol. Cell. Biol. 19: 1202–1209

  • Mahajan R, Delphin C, Guan T, Gerace L, Melchior F . 1997 Cell 88: 97–107

  • Mao Y, Sun M, Desai SD, Liu LF . 2000 Proc. Natl. Acad. Sci. USA 97: 4046–4051

  • Matunis MJ, Coutavas E, Blobel G . 1996 J. Cell Biol. 135: 1457–1470

  • Meluh PB, Koshland D . 1995 Mol. Biol. Cell 6: 793–807

  • Minty A, Dumont X, Kaghad M, Caput D . 2000 J. Biol. Chem. 275: 36316–36323

  • Momand J, Wu HH,, Dasgupta G . 2000 Gene 242: 15–29

  • Muller S, Berger M, Lehembre F, Seeler JS, Haupt Y, Dejean A . 2000 J. Biol. Chem. 275: 13321–13329

  • Muller S, Matunis MJ, Dejean A . 1998 EMBO J. 17: 61–70

  • Okuma T, Honda R, Ichikawa G, Tsumagari N, Yasuda H . 1999 Biochem. Biophys. Res. Commun. 254: 693–698

  • Okura T, Gong L, Kamitani T, Wada T, Okura I, Wei CF, Chang HM, Yeh ET . 1996 J. Immunol. 157: 4277–4281

  • Piette J, Neel H, Marechal V . 1997 Oncogene 15: 1001–1010

  • Reimer G, Pollard KM, Penning CA, Ochs RL, Lischwe MA, Busch H, Tan EM . 1987 Arthritis Rheum. 30: 793–800

  • Rodriguez MS, Desterro JM, Lain S, Midgley CA, Lane DP, Hay RT . 1999 EMBO J. 18: 6455–6461

  • Roth J, Dobbelstein M, Freedman DA, Shenk T, Levine AJ . 1998 EMBO J. 17: 554–564

  • Rubbi CP, Milner J . 2000 Oncogene 19: 85–96

  • Saitoh H, Pu RT, Dasso M . 1997 Trends Biochem. Sci. 10: 374–376

  • Shaulsky G, Goldfinger N, Ben-Ze'ev A, Rotter V . 1990 Mol. Cell. Biol. 10: 6565–6577

  • Shen Z, Pardington-Purtymun PE, Comeaux JC, Moyzis RK, Chen DJ . 1996 Genomics 37: 183–186

  • Shieh SY, Ahn J, Tamai K, Taya Y, Prives C . 2000 Genes Dev. 14: 289–300

  • Sionov RV, Haupt Y . 1999 Oncogene 18: 6145–6157

  • Snow CM, Senior A, Gerace L . 1987 J. Cell Biol. 104: 1143–1156

  • Sternsdorf T, Jensen K, Will H . 1997 J. Cell Biol. 139: 1621–1634

  • Sternsdorf T, Jensen K, Reich B, Will H . 1999 J. Biol. Chem. 274: 12555–12566

  • Suzuki T, Ichiyama A, Saitoh H, Kawakami T, Omata M, Chung CH, Kimura M, Shimbara N, Tanaka K . 1999 J. Biol. Chem. 274: 31131–31134

  • Unger T, Juven-Gershon T, Moallem E, Berger M, Vogt Sionov R, Lozano G, Oren M, Haupt Y . 1999 EMBO J. 18: 1805–1814

  • Waterman MJ, Stavridi ES, Waterman JL, Halazonetis TD . 1998 Nat. Genet. 19: 175–178

  • Zhang Y, Xiong Y . 1999 Mol. Cell. 3: 579–591

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Acknowledgements

We thank Li-Chung Hsu, Zaklina Strezoska, Mikhail Nikiforov and Jason Hill for helpful discussions, Roman Kondratov and Andrei Budanov for technical advice. We thank Hongming Zhu for help with flow cytometry and Meiling Chen for help with confocal microscopy. This work was supported by NIH grant CA75179 to Andrei V Gudkov and NIH NIEHS grant ES08353 to Zhiyuan Shen.

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  1. Zhiyuan Shen

    Present address: Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, 915 Camino de Salud, NE Albuqierque, 871312, NM, USA

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  1. Department of Molecular Genetics, College of Medicine, University of Illinois at Chicago, 900 South Ashland Avenue, Chicago, 60607, Illinois, IL, USA

    Serena SS Kwek, Jason Derry, Angela L Tyner, Zhiyuan Shen & Andrei V Gudkov

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Kwek, S., Derry, J., Tyner, A. et al. Functional analysis and intracellular localization of p53 modified by SUMO-1. Oncogene 20, 2587–2599 (2001). https://doi.org/10.1038/sj.onc.1204362

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