Advertisement

The role of acetylation sites in the regulation of p53 activity

  • Yun Wang
  • Yaqi Chen
  • Qiang Chen
  • Xiuyuan Zhang
  • Hongye Wang
  • Zhonghua WangEmail author
  • Jian WangEmail author
  • Chunyan TianEmail author
Original Article

Abstract

As a “genomic guardian”, p53 mainly functions as a transcription factor that regulates downstream targets responsible for cell fate control, and the activity of p53 is tightly regulated by a complex network that include an abundance of post-translational modifications. Notably, acetylation of p53 at many positions has been demonstrated to play a major role in accurate p53 regulation and cell fate determination. However, no evidence has been provided to compare the effect of acetylation at different sites on p53 regulation. Here, we constructed six acetylation-defective p53 mutants that lysine was substituted by arginine at residues 120, 164, 305, 320, 370/372/373 or 381/382/386, respectively, and determined their effects on p53 activity systematically. Our results showed that all six mutants exhibited diminished transactivation ability and selective regulation of target genes expression through distinct mechanisms. Specifically, lysine 370/372/373 and 381/382/386 mutations decreased p53 stability, and lysine 305 mutation reduced p53 phosphorylation level at serine 15, while lysine 120 and 164 mutations decreased p53 acetylation level at lysine 382. Collectively, these data indicate that acetylation of p53 at different sites has diverse regulatory effects on p53 transcriptional activity through different mechanisms.

Keywords

p53 Acetylation Post-translational modifications Acetylation-defective mutant Regulation 

Abbreviations

PTM

Post-translational modifications

HATs

Histone/lysine acetyltransferases

CTD

C-terminal domain

PCAF

p300/CBP associated factor

K120

Lysine 120

K164

Lysine 164

K305

Lysine 305

K320

Lysine 320

K382/383

Lysine 382/383

S15

Serine 15

pG13-Luc

pG13L containing 13 tandem p53 binding site repeats

K120R

Mutants with lysine to arginine changes at residues 120

K164R

Mutants with lysine to arginine changes at residues 164

K305R

Mutants with lysine to arginine changes at residues 305

K320R

Mutants with lysine to arginine changes at residues 320

K370/372/373R

Mutants with lysine to arginine changes at residues 370/372/373

K381/382/386R

Mutants with lysine to arginine changes at residues 381/382/386

CHX

Cycloheximide

Notes

Acknowledgements

We thank Dr. Bert Vogelstein (Johns Hopkins University) for providing the luciferase reporter plasmid pG13-Luc, Dr. Lingqiang Zhang (Beijing Institute of Lifeomics) for Human lung adenocarcinoma H1299 cell line.

Author contributions

C.T. and J.W. conceived the project and designed the experiments. The experiments were performed by Y.W., Y.C., X.Z., Q.C., and H.W. Data were analyzed by C.T., J.W., Y.C. and Z.W. C.T. wrote the manuscript. Z.W. and J.W. provided critical proof reading of the manuscript.

Funding

This work was partially supported by grants from the National Natural Science Foundation Projects (31270799, 31771563), the National Key Research and Development Program of China (2017YFA0505700), the Chinese Program of International S&T Cooperation (2014DFB30020), and National Modern Agro (Dairy) Industry and Technology System (CARS-37).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Kastenhuber ER, Lowe SW (2017) Putting p53 in context. Cell 170:2017CrossRefGoogle Scholar
  2. 2.
    Mello SS, Attardi LD (2018) Deciphering p53 signaling in tumor suppression. Curr Opin Cell Biol 51:2018CrossRefGoogle Scholar
  3. 3.
    Kruse JP, Gu W (2009) Modes of p53 regulation. Cell 137:2009CrossRefGoogle Scholar
  4. 4.
    Gu B, Zhu WG (2012) Surf the post-translational modification network of p53 regulation. Int J Biol Sci 8:2012CrossRefGoogle Scholar
  5. 5.
    Reed SM, Quelle DE (2014) p53 Acetylation: regulation and consequences. Cancers 7:2014CrossRefGoogle Scholar
  6. 6.
    Zhu WG (2017) Regulation of p53 acetylation. Science China. Life Sci 60:2017Google Scholar
  7. 7.
    Wang SJ, Li D, Ou Y, Jiang L, Chen Y, Zhao Y, Gu W (2016) Acetylation is crucial for p53-mediated ferroptosis and tumor suppression. Cell Rep 17:2016Google Scholar
  8. 8.
    Wang D, Kon N, Lasso G, Jiang L, Leng W, Zhu WG, Qin J, Honig B, Gu W (2016) Acetylation-regulated interaction between p53 and SET reveals a widespread regulatory mode. Nature 538:2016Google Scholar
  9. 9.
    Gu W, Roeder RG (1997) Activation of p53 sequence-specific DNA binding by acetylation of the p53 C-terminal domain. Cell 90:1997CrossRefGoogle Scholar
  10. 10.
    Feng L, Lin T, Uranishi H, Gu W, Xu Y (2005) Functional analysis of the roles of posttranslational modifications at the p53 C terminus in regulating p53 stability and activity. Mol Cell Biol 25:2005Google Scholar
  11. 11.
    Zhao Y, Lu S, Wu L, Chai G, Wang H, Chen Y, Sun J, Yu Y, Zhou W, Zheng Q, Wu M, Otterson GA, Zhu WG (2006) Acetylation of p53 at lysine 373/382 by the histone deacetylase inhibitor depsipeptide induces expression of p21(Waf1/Cip1). Mol Cell Biol 26:2006Google Scholar
  12. 12.
    Tang Y, Zhao W, Chen Y, Zhao Y, Gu W (2008) Acetylation is indispensable for p53 activation. Cell 133:2008CrossRefGoogle Scholar
  13. 13.
    Wang YH, Tsay YG, Tan BC, Lo WY, Lee SC (2003) Identification and characterization of a novel p300-mediated p53 acetylation site, lysine 305. J Biol Chem 278:2003Google Scholar
  14. 14.
    Barlev NA, Liu L, Chehab NH, Mansfield K, Harris KG, Halazonetis TD, Berger SL (2001) Acetylation of p53 activates transcription through recruitment of coactivators/histone acetyltransferases. Mol Cell 8:2001CrossRefGoogle Scholar
  15. 15.
    Knights CD, Catania J, Di Giovanni S, Muratoglu S, Perez R, Swartzbeck A, Quong AA, Zhang X, Beerman T, Pestell RG, Avantaggiati ML (2006) Distinct p53 acetylation cassettes differentially influence gene-expression patterns and cell fate. J Cell Biol 173:2006CrossRefGoogle Scholar
  16. 16.
    Sykes SM, Mellert HS, Holbert MA, Li K, Marmorstein R, Lane WS, McMahon SB (2006) Acetylation of the p53 DNA-binding domain regulates apoptosis induction. Mol Cell 24:2006CrossRefGoogle Scholar
  17. 17.
    Tang Y, Luo J, Zhang W, Gu W (2006) Tip60-dependent acetylation of p53 modulates the decision between cell-cycle arrest and apoptosis. Mol Cell 24:2006CrossRefGoogle Scholar
  18. 18.
    Wang S, Tian C, Xiao T, Xing G, He F, Zhang L, Chen H (2010) Differential regulation of Apak by various DNA damage signals. Mol Cell Biochem 333:2010CrossRefGoogle Scholar
  19. 19.
    Tian C, Xing G, Xie P, Lu K, Nie J, Wang J, Li L, Gao M, Zhang L, He F (2009) KRAB-type zinc-finger protein Apak specifically regulates p53-dependent apoptosis. Nat Cell Biol 11:2009Google Scholar
  20. 20.
    Wang SJ, Gu W (2014) To be, or not to be: functional dilemma of p53 metabolic regulation. Curr Opin Oncol 26:2014Google Scholar
  21. 21.
    Menendez D, Inga A, Resnick MA (2009) The expanding universe of p53 targets. Nat Rev Cancer 9:2009CrossRefGoogle Scholar
  22. 22.
    Shu KX, Li B, Wu LX (2007) The p53 network: p53 and its downstream genes. Colloids Surf B 55:2007CrossRefGoogle Scholar
  23. 23.
    Li M, Luo J, Brooks CL, Gu W (2002) Acetylation of p53 inhibits its ubiquitination by Mdm2. J Biol Chem 277:2002Google Scholar
  24. 24.
    Ito A, Kawaguchi Y, Lai CH, Kovacs JJ, Higashimoto Y, Appella E, Yao TP (2002) MDM2-HDAC1-mediated deacetylation of p53 is required for its degradation. EMBO J 21:2002Google Scholar
  25. 25.
    Meulmeester E, Pereg Y, Shiloh Y, Jochemsen AG (2005) ATM-mediated phosphorylations inhibit Mdmx/Mdm2 stabilization by HAUSP in favor of p53 activation. Cell Cycle 4:2005CrossRefGoogle Scholar
  26. 26.
    Loughery J, Cox M, Smith LM, Meek DW (2014) Critical role for p53-serine 15 phosphorylation in stimulating transactivation at p53-responsive promoters. Nucleic Acids Res 42:2014CrossRefGoogle Scholar
  27. 27.
    Chao C, Wu Z, Mazur SJ, Borges H, Rossi M, Lin T, Wang JY, Anderson CW, Appella E, Xu Y (2006) Acetylation of mouse p53 at lysine 317 negatively regulates p53 apoptotic activities after DNA damage. Mol Cell Biol 26:2006CrossRefGoogle Scholar
  28. 28.
    Rodriguez MS, Desterro JM, Lain S, Lane DP, Hay RT (2000) Multiple C-terminal lysine residues target p53 for ubiquitin-proteasome-mediated degradation. Mol Cell Biol 20:2000Google Scholar
  29. 29.
    Nakamura S, Roth JA, Mukhopadhyay T (2000) Multiple lysine mutations in the C-terminal domain of p53 interfere with MDM2-dependent protein degradation and ubiquitination. Mol Cell Biol 20:2000Google Scholar
  30. 30.
    Ou YH, Chung PH, Sun TP, Shieh SY (2005) p53 C-terminal phosphorylation by CHK1 and CHK2 participates in the regulation of DNA-damage-induced C-terminal acetylation. Mol Biol Cell 16:2005CrossRefGoogle Scholar
  31. 31.
    Jenkins LM, Yamaguchi H, Hayashi R, Cherry S, Tropea JE, Miller M, Wlodawer A, Appella E, Mazur SJ (2009) Two distinct motifs within the p53 transactivation domain bind to the Taz2 domain of p300 and are differentially affected by phosphorylation. Biochemistry 48:2009CrossRefGoogle Scholar
  32. 32.
    Lee CW, Ferreon JC, Ferreon AC, Arai M, Wright PE (2010) Graded enhancement of p53 binding to CREB-binding protein (CBP) by multisite phosphorylation. Proc Natl Acad Sci USA 107:2010Google Scholar
  33. 33.
    Lau AW, Liu P, Inuzuka H, Gao D (2014) SIRT1 phosphorylation by AMP-activated protein kinase regulates p53 acetylation. Am J Cancer Res 4:2014Google Scholar
  34. 34.
    Bode AM, Dong Z (2004) Post-translational modification of p53 in tumorigenesis. Nat Rev Cancer 4:2004CrossRefGoogle Scholar
  35. 35.
    Shahar OD, Gabizon R, Feine O, Alhadeff R, Ganoth A, Argaman L, Shimshoni E, Friedler A, Goldberg M (2013) Acetylation of lysine 382 and phosphorylation of serine 392 in p53 modulate the interaction between p53 and MDC1 in vitro. PLoS ONE 8:2013CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.College of Animal ScienceShandong Agricultural UniversityTai’anChina
  2. 2.State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing)Beijing Institute of LifeomicsBeijingChina
  3. 3.School of Public HealthShandong First Medical University & Shandong Academy of Medical ScienceTai’anChina

Personalised recommendations