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Stress-induced changes in CARF expression determine cell fate to death, survival, or malignant transformation

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Cell Stress and Chaperones Aims and scope

Abstract

CARF (Collaborator of ARF) was discovered as an ARF-interacting protein that activated ARF-p53-p21WAF1 signaling involved in cellular response to a variety of stresses, including oxidative, genotoxic, oncogenic, or telomere deprotection stresses, leading to senescence, growth arrest, or apoptosis. Of note, whereas suppression of CARF was lethal, its enrichment was associated with increased proliferation and malignant transformation of cells. These reports have predicted that CARF could serve as a multi-stress marker with a predictive value for cell fates. Here, we recruited various in vitro stress models and examined their effect on CARF expression using human normal fibroblasts. We demonstrate that CARF levels in stress and post-stress conditions could predict the fate of cells towards either death or enhanced proliferation and malignant transformation. We provide extensive molecular evidence that (i) CARF expression changes in response to stress, (ii) it modulates cell death or survival signaling and determines the fate of cells, and (iii) it may serve as a predictive measure of cellular response to stress and an important marker for biosafety.

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Abbreviations

CARF:

Collaborator of ARF

DDR:

DNA damage response

UPR:

Unfolded protein response

ROS:

Reactive oxygen species

CTA:

Cell transformation assay

IC:

Inhibitory concentration

ER:

Endoplasmic reticulum

References

  • Adams PD, Jasper H, Rudolph KL (2015) Aging-induced stem cell mutations as drivers for disease and cancer. Cell Stem Cell 16:601–612

    Article  CAS  Google Scholar 

  • Armstrong JL, Flockhart R, Veal GJ, Lovat PE, Redfern CP (2010) Regulation of endoplasmic reticulum stress-induced cell death by ATF4 in neuroectodermal tumor cells. J Biol Chem 285:6091–6100

    Article  CAS  Google Scholar 

  • Balducci L, Ershler WB (2005) Cancer and ageing: a nexus at several levels. Nat Rev Cancer 5:655–662

    Article  CAS  Google Scholar 

  • Bendjennat M, Boulaire J, Jascur T, Brickner H, Barbier V, Sarasin A, Fotedar A, Fotedar R (2003) UV irradiation triggers ubiquitin-dependent degradation of p21(WAF1) to promote DNA repair. Cell 114:599–610

    Article  CAS  Google Scholar 

  • Benhar M, Engelberg D, Levitzki A (2002) ROS, stress-activated kinases and stress signaling in cancer. EMBO Rep 3:420–425

    Article  CAS  Google Scholar 

  • Benz CC, Yau C (2008) Ageing, oxidative stress and cancer: paradigms in parallax. Nat Rev Cancer 8:875–879

    Article  CAS  Google Scholar 

  • Bhagat L, Singh VP, Dawra RK, Saluja AK (2008) Sodium arsenite induces heat shock protein 70 expression and protects against secretagogue-induced trypsinogen and NF-kappaB activation. J Cell Physiol 215:37–46

    Article  CAS  Google Scholar 

  • Boudreault S, Martenon-Brodeur C, Caron M, Garant JM, Tremblay MP, Armero VE, Durand M, Lapointe E, Thibault P, Tremblay-Letourneau M, Perreault JP, Scott MS, Lemay G, Bisaillon M (2016) Global profiling of the cellular alternative RNA splicing landscape during virus-host interactions. PLoS One 11:e0161914

    Article  Google Scholar 

  • Cheung CT, Singh R, Yoon AR, Hasan MK, Yaguchi T, Kaul SC, Yun CO, Wadhwa R (2011) Molecular characterization of apoptosis induced by CARF silencing in human cancer cells. Cell Death Differ 18:589–601

  • Cheung CT, Hasan MK, Widodo N, Kaul SC, Wadhwa R (2009) CARF: an emerging regulator of p53 tumor suppressor and senescence pathway. Mech Ageing Dev 130:18–23

    Article  CAS  Google Scholar 

  • Cheung CT, Kaul SC, Wadhwa R (2010) Molecular bridging of aging and cancer: a CARF link. Ann N Y Acad Sci 1197:129–133

    Article  CAS  Google Scholar 

  • Cheung CT, Singh R, Kalra RS, Kaul SC, Wadhwa R (2014) Collaborator of ARF (CARF) regulates proliferative fate of human cells by dose-dependent regulation of DNA damage signaling. J Biol Chem 289:18258–18269

    Article  CAS  Google Scholar 

  • Evans CG, Chang L, Gestwicki JE (2010) Heat shock protein 70 (hsp70) as an emerging drug target. J Med Chem 53:4585–4602

    Article  CAS  Google Scholar 

  • Fullove TP, Yu H (2013) DNA damage and repair of human skin keratinocytes concurrently exposed to pyrene derivatives and UVA light. Toxicol Res (Camb) 2:193–199

    Article  CAS  Google Scholar 

  • Hasan MK, Yaguchi T, Sugihara T, Kumar PK, Taira K, Reddel RR, Kaul SC, Wadhwa R (2002) CARF is a novel protein that cooperates with mouse p19ARF (human p14ARF) in activating p53. J Biol Chem 277:37765–37770

    Article  CAS  Google Scholar 

  • Hasan MK, Yaguchi T, Minoda Y, Hirano T, Taira K, Wadhwa R, Kaul SC (2004) Alternative reading frame protein (ARF)-independent function of CARF (collaborator of ARF) involves its interactions with p53: evidence for a novel p53-activation pathway and its negative feedback control. Biochem J 380:605–610

    Article  CAS  Google Scholar 

  • Hasan MK, Wadhwa R, Kaul SC (2007) CARF binds to three members (ARF, p53, and HDM2) of the p53 tumor-suppressor pathway. Ann N Y Acad Sci 1100:312–315

    Article  Google Scholar 

  • Hasan MK, Yaguchi T, Harada JI, Hirano T, Wadhwa R, Kaul SC (2008) CARF (collaborator of ARF) interacts with HDM2: evidence for a novel regulatory feedback regulation of CARF-p53-HDM2-p21WAF1 pathway. Int J Oncol 32:663–671

    CAS  PubMed  Google Scholar 

  • Hasan K, Cheung C, Kaul Z, Shah N, Sakaushi S, Sugimoto K, Oka S, Kaul SC, Wadhwa R (2009) CARF is a vital dual regulator of cellular senescence and apoptosis. J Biol Chem 284:1664–1672

    Article  CAS  Google Scholar 

  • Hayashi K, Sasaki K, Asada S, Tsuchiya T, Hayashi M, Yoshimura I, Tanaka N, Umeda M (2008) Technical modification of the Balb/c 3T3 cell transformation assay: the use of serum-reduced medium to optimise the practicability of the protocol. Altern Lab Anim 36:653–665

    Article  CAS  Google Scholar 

  • Huang HL, Wu YC, Su LJ, Huang YJ, Charoenkwan P, Chen WL, Lee HC, Chu WC, Ho SY (2015) Discovery of prognostic biomarkers for predicting lung cancer metastasis using microarray and survival data. BMC Bioinformatics 16:54

    Article  Google Scholar 

  • Kalra RS, Cheung CT, Chaudhary A, Prakash J, Kaul SC, Wadhwa R (2015) CARF (collaborator of ARF) overexpression in p53-deficient cells promotes carcinogenesis. Mol Oncol 9:1877–1889

    Article  CAS  Google Scholar 

  • Kalra RS, Chaudhary A, Yoon AR, Bhargava P, Omar A, Garg S, Yun CO, Kaul SC, Wadhwa R (2018) CARF enrichment promotes epithelial-mesenchymal transition via Wnt/beta-catenin signaling: its clinical relevance and potential as a therapeutic target. Oncogenesis 7:39

    Article  Google Scholar 

  • Kondo S, Toyokuni S, Iwasa Y, Tanaka T, Onodera H, Hiai H, Imamura M (1999) Persistent oxidative stress in human colorectal carcinoma, but not in adenoma. Free Radic Biol Med 27:401–410

    Article  CAS  Google Scholar 

  • Krtolica A, Parrinello S, Lockett S, Desprez PY, Campisi J (2001) Senescent fibroblasts promote epithelial cell growth and tumorigenesis: a link between cancer and aging. Proc Natl Acad Sci U S A 98:12072–12077

    Article  CAS  Google Scholar 

  • Mansilla SF, Soria G, Vallerga MB, Habif M, Martinez-Lopez W, Prives C, Gottifredi V (2013) UV-triggered p21 degradation facilitates damaged-DNA replication and preserves genomic stability. Nucleic Acids Res 41:6942–6951

    Article  CAS  Google Scholar 

  • Miki TS, Richter H, Ruegger S, Grosshans H (2014) PAXT-1 promotes XRN2 activity by stabilizing it through a conserved domain. Mol Cell 53:351–360

    Article  CAS  Google Scholar 

  • Mosser DD, Caron AW, Bourget L, Denis-Larose C, Massie B (1997) Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis. Mol Cell Biol 17:5317–5327

    Article  CAS  Google Scholar 

  • Nuntharatanapong N, Chen K, Sinhaseni P, Keaney JF Jr (2005) EGF receptor-dependent JNK activation is involved in arsenite-induced p21Cip1/Waf1 upregulation and endothelial apoptosis. Am J Physiol Heart Circ Physiol 289:H99–H107

    Article  CAS  Google Scholar 

  • Nylandsted J, Gyrd-Hansen M, Danielewicz A, Fehrenbacher N, Lademann U, Hoyer-Hansen M, Weber E, Multhoff G, Rohde M, Jaattela M (2004) Heat shock protein 70 promotes cell survival by inhibiting lysosomal membrane permeabilization. J Exp Med 200:425–435

    Article  CAS  Google Scholar 

  • Sato S, Ishikawa H, Yoshikawa H, Izumikawa K, Simpson RJ, Takahashi N (2015) Collaborator of alternative reading frame protein (CARF) regulates early processing of pre-ribosomal RNA by retaining XRN2 (5′-3′ exoribonuclease) in the nucleoplasm. Nucleic Acids Res 43:10397–10410

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sau A, Pellizzari Tregno F, Valentino F, Federici G, Caccuri AM (2010) Glutathione transferases and development of new principles to overcome drug resistance. Arch Biochem Biophys 500:116–122

    Article  CAS  Google Scholar 

  • Shan J, Fu L, Balasubramanian MN, Anthony T, Kilberg MS (2012) ATF4-dependent regulation of the JMJD3 gene during amino acid deprivation can be rescued in Atf4-deficient cells by inhibition of deacetylation. J Biol Chem 287:36393–36403

    Article  CAS  Google Scholar 

  • Singh R, Kalra RS, Hasan K, Kaul Z, Cheung CT, Huschtscha L, Reddel RR, Kaul SC, Wadhwa R (2014) Molecular characterization of collaborator of ARF (CARF) as a DNA damage response and cell cycle checkpoint regulatory protein. Exp Cell Res 322:324–334

    Article  CAS  Google Scholar 

  • Takabe W, Niki E, Uchida K, Yamada S, Satoh K, Noguchi N (2001) Oxidative stress promotes the development of transformation: involvement of a potent mutagenic lipid peroxidation product, acrolein. Carcinogenesis 22:935–941

    Article  CAS  Google Scholar 

  • Tew KD, Townsend DM (2012) Glutathione-s-transferases as determinants of cell survival and death. Antioxid Redox Signal 17:1728–1737

    Article  CAS  Google Scholar 

  • Tsalikis J, Pan Q, Tattoli I, Maisonneuve C, Blencowe BJ, Philpott DJ, Girardin SE (2016) The transcriptional and splicing landscape of intestinal organoids undergoing nutrient starvation or endoplasmic reticulum stress. BMC Genomics 17:680

    Article  Google Scholar 

  • Wortel IMN, van der Meer LT, Kilberg MS, van Leeuwen FN (2017) Surviving stress: modulation of ATF4-mediated stress responses in normal and malignant cells. Trends Endocrinol Metab 28:794–806

    Article  CAS  Google Scholar 

  • Yang CS, Chang KY, Rana TM (2014) Genome-wide functional analysis reveals factors needed at the transition steps of induced reprogramming. Cell Rep 8:327–337

    Article  CAS  Google Scholar 

  • Zhang H, Reynolds M (2019) Cadmium exposure in living organisms: a short review. Sci Total Environ 678:761–767

    Article  CAS  Google Scholar 

Download references

Funding

The present study is supported by the grants from the Department of Biotechnology (Government of India) and AIST (Japan).

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Authors and Affiliations

  1. AIST-INDIA DAILAB, National Institute of Advanced Industrial Science & Technology (AIST), Tsukuba, Ibaraki, 305-8565, Japan

    Rajkumar S. Kalra, Anupama Chaudhary, Amr Omar, Caroline T. Cheung, Sukant Garg, Sunil C. Kaul & Renu Wadhwa

Authors
  1. Rajkumar S. Kalra
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  2. Anupama Chaudhary
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  3. Amr Omar
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  4. Caroline T. Cheung
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  5. Sukant Garg
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  6. Sunil C. Kaul
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  7. Renu Wadhwa
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Contributions

RSK, RW, and SCK designed the study, interpreted the results, and wrote the manuscript. RSK, AC, AO, and SG performed the experiments and coordinated in result compilation. CTC helped in interpretation of results and manuscript writing.

Corresponding authors

Correspondence to Sunil C. Kaul or Renu Wadhwa.

Ethics declarations

All experiments were performed according to the approval by institute ethical committee.

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The authors declare that they have no conflict of interest.

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Kalra, R.S., Chaudhary, A., Omar, A. et al. Stress-induced changes in CARF expression determine cell fate to death, survival, or malignant transformation. Cell Stress and Chaperones 25, 481–494 (2020). https://doi.org/10.1007/s12192-020-01088-y

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  • DOI: https://doi.org/10.1007/s12192-020-01088-y

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