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The role of acetylation sites in the regulation of p53 activity

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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.

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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

References

  1. Kastenhuber ER, Lowe SW (2017) Putting p53 in context. Cell 170:2017

    Article  Google Scholar 

  2. Mello SS, Attardi LD (2018) Deciphering p53 signaling in tumor suppression. Curr Opin Cell Biol 51:2018

    Article  Google Scholar 

  3. Kruse JP, Gu W (2009) Modes of p53 regulation. Cell 137:2009

    Article  Google Scholar 

  4. Gu B, Zhu WG (2012) Surf the post-translational modification network of p53 regulation. Int J Biol Sci 8:2012

    Article  Google Scholar 

  5. Reed SM, Quelle DE (2014) p53 Acetylation: regulation and consequences. Cancers 7:2014

    Article  Google Scholar 

  6. Zhu WG (2017) Regulation of p53 acetylation. Science China. Life Sci 60:2017

    Google Scholar 

  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 17:2016

    Google Scholar 

  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 538:2016

    Google Scholar 

  9. Gu W, Roeder RG (1997) Activation of p53 sequence-specific DNA binding by acetylation of the p53 C-terminal domain. Cell 90:1997

    Article  Google Scholar 

  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 25:2005

    Google Scholar 

  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 26:2006

    Google Scholar 

  12. Tang Y, Zhao W, Chen Y, Zhao Y, Gu W (2008) Acetylation is indispensable for p53 activation. Cell 133:2008

    Article  Google Scholar 

  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 278:2003

    Google Scholar 

  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 8:2001

    Article  Google Scholar 

  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 173:2006

    Article  Google Scholar 

  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 24:2006

    Article  Google Scholar 

  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 24:2006

    Article  Google Scholar 

  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 333:2010

    Article  Google Scholar 

  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 11:2009

    Google Scholar 

  20. Wang SJ, Gu W (2014) To be, or not to be: functional dilemma of p53 metabolic regulation. Curr Opin Oncol 26:2014

    Google Scholar 

  21. Menendez D, Inga A, Resnick MA (2009) The expanding universe of p53 targets. Nat Rev Cancer 9:2009

    Article  Google Scholar 

  22. Shu KX, Li B, Wu LX (2007) The p53 network: p53 and its downstream genes. Colloids Surf B 55:2007

    Article  Google Scholar 

  23. Li M, Luo J, Brooks CL, Gu W (2002) Acetylation of p53 inhibits its ubiquitination by Mdm2. J Biol Chem 277:2002

    Google Scholar 

  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 21:2002

    Google Scholar 

  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 4:2005

    Article  Google Scholar 

  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 42:2014

    Article  Google Scholar 

  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 26:2006

    Article  Google Scholar 

  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 20:2000

    Google Scholar 

  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 20:2000

    Google Scholar 

  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 16:2005

    Article  Google Scholar 

  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 48:2009

    Article  Google Scholar 

  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 107:2010

    Google Scholar 

  33. Lau AW, Liu P, Inuzuka H, Gao D (2014) SIRT1 phosphorylation by AMP-activated protein kinase regulates p53 acetylation. Am J Cancer Res 4:2014

    Google Scholar 

  34. Bode AM, Dong Z (2004) Post-translational modification of p53 in tumorigenesis. Nat Rev Cancer 4:2004

    Article  Google Scholar 

  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 8:2013

    Article  Google Scholar 

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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.

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).

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Author notes

  1. Yun Wang, Yaqi Chen and Qiang Chen contributed equally to this work.

Authors and Affiliations

  1. College of Animal Science, Shandong Agricultural University, Tai’an, Shandong Province, 271018, China

    Yun Wang & Zhonghua Wang

  2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, 102206, China

    Yaqi Chen, Xiuyuan Zhang, Hongye Wang, Jian Wang & Chunyan Tian

  3. School of Public Health, Shandong First Medical University & Shandong Academy of Medical Science, Tai’an, Shandong Province, 271016, China

    Qiang Chen

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  1. Yun Wang
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  4. Xiuyuan Zhang
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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.

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Correspondence to Zhonghua Wang, Jian Wang or Chunyan Tian.

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Wang, Y., Chen, Y., Chen, Q. et al. The role of acetylation sites in the regulation of p53 activity. Mol Biol Rep 47, 381–391 (2020). https://doi.org/10.1007/s11033-019-05141-7

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  • DOI: https://doi.org/10.1007/s11033-019-05141-7

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