Abstract
Protein kinase B (PKB/Akt) is ubiquitously expressed in cells. Phosphorylation of its multiple targets in response to various stimuli, including growth factors or cytokines, promotes cell survival and inhibits apoptosis. PKB is upregulated in many different cancers and a significant amount of the enzyme is present in its activated form. Here we show that PKB phosphorylates one of the anti-apoptotic proteins—transcription factor Twist-1 at Ser42. Cells expressing Twist-1 displayed inefficient p53 upregulation in response to DNA damage induced by γ-irradiation or the genotoxic drug adriamycin. This influenced the activation of p53 target genes such as p21Waf1 and Bax and led to aberrant cell-cycle regulation and the inhibition of apoptosis. The impaired induction of these p53 effector molecules is likely to be mediated by PKB-dependent phosphorylation of Twist-1 because, unlike the wild-type mutant, the Twist-1 S42A mutant did not confer cell resistance to DNA damage. Moreover, phosphorylation of Twist-1 at Ser42 was shown in vivo in various human cancer tissues, suggesting that this post-translational modification ensures functional activation of Twist-1 after promotion of survival during carcinogenesis.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ansieau S, Bastid J, Doreau A, Morel AP, Bouchet BP, Thomas C et al. (2008). Induction of EMT by twist proteins as a collateral effect of tumor-promoting inactivation of premature senescence. Cancer Cell 14: 79–89.
Bialek P, Kern B, Yang X, Schrock M, Sosic D, Hong N et al. (2004). A twist code determines the onset of osteoblast differentiation. Dev Cell 6: 423–435.
Biggs WH, Meisenhelder J, Hunter T, Cavenee WK, Arden KC . (1999). Protein kinase B/Akt-mediated phosphorylation promotes nuclear exclusion of the winged helix transcription factor FKHR1. Proc Natl Acad Sci USA 96: 7421–7426.
Blom N, Sicheritz-Ponten T, Gupta R, Gammeltoft S, Brunak S . (2004). Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. Proteomics 4: 1633–1649.
Bozulic L, Surucu B, Hynx D, Hemmings BA . (2008). PKBalpha/Akt1 acts downstream of DNA-PK in the DNA double-strand break response and promotes survival. Mol Cell 30: 203–213.
Cai J, Jabs EW . (2005). A twisted hand: bHLH protein phosphorylation and dimerization regulate limb development. Bioessays 27: 1102–1106.
Carpten JD, Faber AL, Horn C, Donoho GP, Briggs SL, Robbins CM et al. (2007). A transforming mutation in the pleckstrin homology domain of AKT1 in cancer. Nature 448: 439–444.
Cheng GZ, Chan J, Wang Q, Zhang W, Sun CD, Wang LH . (2007). Twist transcriptionally up-regulates AKT2 in breast cancer cells leading to increased migration, invasion, and resistance to paclitaxel. Cancer Res 67: 1979–1987.
Di Cristofano A, Pesce B, Cordon-Cardo C, Pandolfi PP . (1998). Pten is essential for embryonic development and tumour suppression. Nat Genet 19: 348–355.
el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM et al. (1993). WAF1, a potential mediator of p53 tumor suppression. Cell 75: 817–825.
Elias MC, Tozer KR, Silber JR, Mikheeva S, Deng M, Morrison RS et al. (2005). TWIST is expressed in human gliomas and promotes invasion. Neoplasia 7: 824–837.
Fayard E, Tintignac LA, Baudry A, Hemmings BA . (2005). Protein kinase B/Akt at a glance. J Cell Sci 118: 5675–5678.
Feng J, Tamaskovic R, Yang Z, Brazil DP, Merlo A, Hess D et al. (2004). Stabilization of Mdm2 via decreased ubiquitination is mediated by protein kinase B/Akt-dependent phosphorylation. J Biol Chem 279: 35510–35517.
Firulli AB, Conway SJ . (2008). Phosphoregulation of Twist1 provides a mechanism of cell fate control. Curr Med Chem 15: 2641–2647.
Firulli BA, Krawchuk D, Centonze VE, Vargesson N, Virshup DM, Conway SJ et al. (2005). Altered Twist1 and Hand2 dimerization is associated with Saethre-Chotzen syndrome and limb abnormalities. Nat Genet 37: 373–381.
Fogal V, Gostissa M, Sandy P, Zacchi P, Sternsdorf T, Jensen K et al. (2000). Regulation of p53 activity in nuclear bodies by a specific PML isoform. EMBO J 19: 6185–6195.
Franke TF . (2008). PI3K/Akt: getting it right matters. Oncogene 27: 6473–6488.
Funato N, Twigg SR, Higashihori N, Ohyama K, Wall SA, Wilkie AO et al. (2005). Functional analysis of natural mutations in two TWIST protein motifs. Hum Mutat 25: 550–556.
Ghouzzi V, Legeai-Mallet L, Benoist-Lasselin C, Lajeunie E, Renier D, Munnich A et al. (2001). Mutations in the basic domain and the loop-helix II junction of TWIST abolish DNA binding in Saethre-Chotzen syndrome. FEBS Lett 492: 112–118.
Gitelman I . (1997). Twist protein in mouse embryogenesis. Dev Biol 189: 205–214.
Gripp KW, Zackai EH, Stolle CA . (2000). Mutations in the human TWIST gene. Hum Mutat 15: 150–155.
Guenou H, Kaabeche K, Dufour C, Miraoui H, Marie PJ . (2006). Down-regulation of ubiquitin ligase Cbl induced by twist haploinsufficiency in Saethre-Chotzen syndrome results in increased PI3K/Akt signaling and osteoblast proliferation. Am J Pathol 169: 1303–1311.
Hamamori Y, Sartorelli V, Ogryzko V, Puri PL, Wu HY, Wang JY et al. (1999). Regulation of histone acetyltransferases p300 and PCAF by the bHLH protein twist and adenoviral oncoprotein E1A. Cell 96: 405–413.
Hayashi M, Nimura K, Kashiwagi K, Harada T, Takaoka K, Kato H et al. (2007). Comparative roles of Twist-1 and Id1 in transcriptional regulation by BMP signaling. J Cell Sci 120: 1350–1357.
Hess D, Keusch JJ, Oberstein SA, Hennekam RC, Hofsteenge J . (2008). Peters Plus syndrome is a new congenital disorder of glycosylation and involves defective Omicron-glycosylation of thrombospondin type 1 repeats. J Biol Chem 283: 7354–7360.
Hosono S, Kajiyama H, Terauchi M, Shibata K, Ino K, Nawa A et al. (2007). Expression of Twist increases the risk for recurrence and for poor survival in epithelial ovarian carcinoma patients. Br J Cancer 96: 314–320.
Kops GJ, de Ruiter ND, Vries-Smits AM, Powell DR, Bos JL, Burgering BM . (1999). Direct control of the Forkhead transcription factor AFX by protein kinase B. Nature 398: 630–634.
Kruse JP, Gu W . (2009). Modes of p53 regulation. Cell 137: 609–622.
Kwok WK, Ling MT, Lee TW, Lau TC, Zhou C, Zhang X et al. (2005). Up-regulation of TWIST in prostate cancer and its implication as a therapeutic target. Cancer Res 65: 5153–5162.
Kwok WK, Ling MT, Yuen HF, Wong YC, Wang X . (2007). Role of p14ARF in TWIST-mediated senescence in prostate epithelial cells. Carcinogenesis 28: 2467–2475.
Kyo S, Sakaguchi J, Ohno S, Mizumoto Y, Maida Y, Hashimoto M et al. (2006). High Twist expression is involved in infiltrative endometrial cancer and affects patient survival. Hum Pathol 37: 431–438.
Liang J, Zubovitz J, Petrocelli T, Kotchetkov R, Connor MK, Han K et al. (2002). PKB/Akt phosphorylates p27, impairs nuclear import of p27 and opposes p27-mediated G1 arrest. Nat Med 8: 1153–1160.
Mironchik Y, Winnard Jr PT, Vesuna F, Kato Y, Wildes F, Pathak AP et al. (2005). Twist overexpression induces in vivo angiogenesis and correlates with chromosomal instability in breast cancer. Cancer Res 65: 10801–10809.
Perkins DN, Pappin DJ, Creasy DM, Cottrell JS . (1999). Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20: 3551–3567.
Pham CG, Bubici C, Zazzeroni F, Knabb JR, Papa S, Kuntzen C et al. (2007). Upregulation of Twist-1 by NF-kappaB blocks cytotoxicity induced by chemotherapeutic drugs. Mol Cell Biol 27: 3920–3935.
Puisieux A, Valsesia-Wittmann S, Ansieau S . (2006). A twist for survival and cancer progression. Br J Cancer 94: 13–17.
Restuccia DF, Hemmings BA . (2009). Cell signaling. Blocking Akt-ivity. Science 325: 1083–1084.
Shibata K, Kajiyama H, Ino K, Terauchi M, Yamamoto E, Nawa A et al. (2008). Twist expression in patients with cervical cancer is associated with poor disease outcome. Ann Oncol 19: 81–85.
Shiota M, Izumi H, Onitsuka T, Miyamoto N, Kashiwagi E, Kidani A et al. (2008). Twist promotes tumor cell growth through YB-1 expression. Cancer Res 68: 98–105.
Singh S, Gramolini AO . (2009). Characterization of sequences in human TWIST required for nuclear localization. BMC Cell Biol 10: 47.
Smit MA, Geiger TR, Song JY, Gitelman I, Peeper DS . (2009). A Twist-Snail axis critical for TrkB-induced epithelial-mesenchymal transition-like transformation, anoikis resistance, and metastasis. Mol Cell Biol 29: 3722–3737.
Sosic D, Richardson JA, Yu K, Ornitz DM, Olson EN . (2003). Twist regulates cytokine gene expression through a negative feedback loop that represses NF-kappaB activity. Cell 112: 169–180.
Spicer DB, Rhee J, Cheung WL, Lassar AB . (1996). Inhibition of myogenic bHLH and MEF2 transcription factors by the bHLH protein Twist. Science 272: 1476–1480.
Stasinopoulos IA, Mironchik Y, Raman A, Wildes F, Winnard Jr P, Raman V . (2005). HOXA5-twist interaction alters p53 homeostasis in breast cancer cells. J Biol Chem 280: 2294–2299.
Thisse B, el Messal M, Perrin-Schmitt F . (1987). The twist gene: isolation of a Drosophila zygotic gene necessary for the establishment of dorsoventral pattern. Nucleic Acids Res 15: 3439–3453.
Valsesia-Wittmann S, Magdeleine M, Dupasquier S, Garin E, Jallas AC, Combaret V et al. (2004). Oncogenic cooperation between H-Twist and N-Myc overrides failsafe programs in cancer cells. Cancer Cell 6: 625–630.
Vichalkovski A, Gresko E, Cornils H, Hergovich A, Schmitz D, Hemmings BA . (2008). NDR kinase is activated by RASSF1A/MST1 in response to Fas receptor stimulation and promotes apoptosis. Curr Biol 18: 1889–1895.
Yang J, Cron P, Thompson V, Good VM, Hess D, Hemmings BA et al. (2002). Molecular mechanism for the regulation of protein kinase B/Akt by hydrophobic motif phosphorylation. Mol Cell 9: 1227–1240.
Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C et al. (2004). Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 117: 927–939.
Yang J, Mani SA, Weinberg RA . (2006). Exploring a new twist on tumor metastasis. Cancer Res 66: 4549–4552.
Yang MH, Wu MZ, Chiou SH, Chen PM, Chang SY, Liu CJ et al. (2008). Direct regulation of TWIST by HIF-1alpha promotes metastasis. Nat Cell Biol 10: 295–305.
Yoeli-Lerner M, Toker A . (2006). Akt/PKB signaling in cancer: a function in cell motility and invasion. Cell Cycle 5: 603–605.
Zhang X, Wang Q, Ling MT, Wong YC, Leung SC, Wang X . (2007). Anti-apoptotic role of TWIST and its association with Akt pathway in mediating taxol resistance in nasopharyngeal carcinoma cells. Int J Cancer 120: 1891–1898.
Zhou BP, Liao Y, Xia W, Spohn B, Lee MH, Hung MC . (2001). Cytoplasmic localization of p21Cip1/WAF1 by Akt-induced phosphorylation in HER-2/neu-overexpressing cells. Nat Cell Biol 3: 245–252.
Acknowledgements
We thank S Bichet and A Bogucki for their help with immunohistochemistry and P King for critical comments on the article. This work was supported by the Novartis Research Foundation and the Swiss Cancer League Grant OCS 01667-02-2005 (to EG).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
Part of the results of this study has been used for the patent application EP 09 174 681.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Rights and permissions
About this article
Cite this article
Vichalkovski, A., Gresko, E., Hess, D. et al. PKB/AKT phosphorylation of the transcription factor Twist-1 at Ser42 inhibits p53 activity in response to DNA damage. Oncogene 29, 3554–3565 (2010). https://doi.org/10.1038/onc.2010.115
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2010.115
Keywords
This article is cited by
-
Role of pelitinib in the regulation of migration and invasion of hepatocellular carcinoma cells via inhibition of Twist1
BMC Cancer (2023)
-
USP13 promotes breast cancer metastasis through FBXL14-induced Twist1 ubiquitination
Cellular Oncology (2023)
-
Norepinephrine promotes glioma cell migration through up-regulating the expression of Twist1
BMC Cancer (2022)
-
Akt: a key transducer in cancer
Journal of Biomedical Science (2022)
-
p32 promotes melanoma progression and metastasis by targeting EMT markers, Akt/PKB pathway, and tumor microenvironment
Cell Death & Disease (2021)