title: Addiction of lung cancer cells to GOF p53 is promoted by up-regulation of epidermal growth factor receptor through multiple contacts with p53 transactivation domain and promoter.
abstract: Human lung cancers harboring gain-of-function (GOF) p53 alleles express higher levels of the epidermal growth factor receptor (EGFR). We demonstrate that a number of GOF p53 alleles directly upregulate EGFR. Knock-down of p53 in lung cancer cells lowers EGFR expression and reduces tumorigenicity and other GOF p53 properties. However, addiction of lung cancer cells to GOF p53 can be compensated by overexpressing EGFR, suggesting that EGFR plays a critical role in addiction. Chromatin immunoprecipitation (ChIP) using lung cancer cells expressing GOF p53 alleles showed that GOF p53 localized to the EGFR promoter. The sequence where GOF p53 is found to interact by ChIP seq can act as a GOF p53 response element. The presence of GOF p53 on the EGFR promoter increased histone H3 acetylation, indicating a mechanism whereby GOF p53 enhances chromatin opening for improved access to transcription factors (TFs). ChIP and ChIP-re-ChIP with p53, Sp1 and CBP histone acetylase (HAT) antibodies revealed docking of GOF p53 on Sp1, leading to increased binding of Sp1 and CBP to the EGFR promoter. Up-regulation of EGFR can occur via GOF p53 contact at other novel sites in the EGFR promoter even when TAD-I is inactivated; these sites are used by both intact and TAD-I mutated GOF p53 and might reflect redundancy in GOF p53 mechanisms for EGFR transactivation. Thus, the oncogenic action of GOF p53 in lung cancer is highly dependent on transactivation of the EGFR promoter via a novel transcriptional mechanism involving coordinated interactions of TFs, HATs and GOF p53.
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Vaughan
authors.affiliation: Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Isabella
authors.initials: I
authors.lastname: Pearsall
authors.affiliation: Integrated Life Sciences Program, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Singh
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.|Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.|Philips Institute, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Brad
authors.initials: B
authors.lastname: Windle
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.|Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.|Integrated Life Sciences Program, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Swati P
authors.initials: SP
authors.lastname: Deb
authors.affiliation: Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.|Division of Hematology, Oncology and Palliative Care, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Grossman
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.|Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.|Philips Institute, Virginia Commonwealth University, Richmond, VA, USA.|Department of Oral Biology, Augusta University, Augusta, GA, USA.
authors.forename: W Andrew
authors.initials: WA
authors.lastname: Yeudall
authors.affiliation: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, VA, USA.|Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA.|Integrated Life Sciences Program, Virginia Commonwealth University, Richmond, VA, USA.
authors.forename: Sumitra
authors.initials: S
authors.lastname: Deb
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D016513:Mice, SCID; D011401:Promoter Regions, Genetic; D016329:Sp1 Transcription Factor / Q000378:metabolism; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D016329:Sp1 Transcription Factor; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D066246:ErbB Receptors
keywords: EGFR; addiction; mutant; p53; transcription
references.citation: Proc Natl Acad Sci U S A. 2010 Jun 1;107(22):10026-31
references.pmid: 20534573
references.citation: J Thorac Dis. 2010 Mar;2(1):48-51
references.pmid: 22263017
references.citation: Mol Cell Biol. 2004 Dec;24(24):10954-64
references.pmid: 15572696
references.citation: Methods Mol Biol. 2009;543:253-66
references.pmid: 19378171
references.citation: Cold Spring Harb Perspect Biol. 2010 Feb;2(2):a001107
references.pmid: 20182618
references.citation: Mol Cell Biol. 2005 Nov;25(22):10097-110
references.pmid: 16260623
references.citation: J Biol Chem. 2001 Oct 19;276(42):39359-67
references.pmid: 11483599
references.citation: Cancer Res. 2000 Dec 15;60(24):6788-93
references.pmid: 11156366
references.citation: Oncogene. 1999 Jan 14;18(2):477-85
references.pmid: 9927204
references.citation: Oncogene. 1994 May;9(5):1341-9
references.pmid: 8152794
references.citation: Mol Cell. 2014 Jun 19;54(6):960-74
references.pmid: 24857548
references.citation: Cell Death Differ. 2013 Apr;20(4):558-66
references.pmid: 23238568
references.citation: Biochemistry. 2013 Aug 27;52(34):5746-59
references.pmid: 23862699
references.citation: Circ Res. 2007 Nov 9;101(10):985-94
references.pmid: 17872466
references.citation: Oncogene. 2003 May 15;22(19):2960-6
references.pmid: 12771947
references.citation: Genes Dev. 2012 Jun 15;26(12):1268-86
references.pmid: 22713868
references.citation: J Biol Chem. 2005 Mar 18;280(11):10047-54
references.pmid: 15647279
references.citation: Nature. 2009 May 7;459(7243):113-7
references.pmid: 19270680
references.citation: FEBS Lett. 2001 Mar 30;493(2-3):65-9
references.pmid: 11286997
references.citation: Oncotarget. 2014 Jan 30;5(2):417-27
references.pmid: 24481480
references.citation: Arch Biochem Biophys. 2012 Feb 1;518(1):79-88
references.pmid: 22198284
references.citation: Oncogene. 2003 Aug 28;22(36):5667-76
references.pmid: 12944915
references.citation: Mol Cell Biol. 1996 Nov;16(11):6009-19
references.pmid: 8887630
references.citation: Oncogene. 1995 Apr 6;10 (7):1275-82
references.pmid: 7731677
references.citation: Oncogene. 1995 Jun 15;10(12):2387-90
references.pmid: 7784087
references.citation: Oncogene. 2003 Oct 20;22(47):7486-95
references.pmid: 14576853
references.citation: Mol Cancer Res. 2009 Aug;7(8):1253-67
references.pmid: 19671680
references.citation: Ann Surg Oncol. 2011 Feb;18(2):543-50
references.pmid: 20811949
references.citation: Cell. 2012 Jan 20;148(1-2):244-58
references.pmid: 22265415
references.citation: Mol Cell Biol. 1998 Apr;18(4):2218-29
references.pmid: 9528793
references.citation: Oncogene. 2006 Jan 19;25(3):359-69
references.pmid: 16170349
references.citation: Cancer Res. 2007 Mar 15;67(6):2396-401
references.pmid: 17363555
references.citation: Oncogene. 1998 Jun 18;16(24):3169-76
references.pmid: 9671396
references.citation: Genes Dev. 2012 Apr 15;26(8):830-45
references.pmid: 22508727
references.citation: Cancer Res. 2006 Aug 1;66(15):7589-97
references.pmid: 16885358
references.citation: Cancer Res. 2004 Oct 15;64(20):7447-54
references.pmid: 15492269
references.citation: Cancer Res. 2007 Feb 15;67(4):1563-70
references.pmid: 17308095
references.citation: Oncogene. 2005 May 19;24(22):3619-31
references.pmid: 15782138
references.citation: Mol Syst Biol. 2005;1:2005.0010
references.pmid: 16729045
references.citation: Genes Cancer. 2011 Oct;2(10):943-55
references.pmid: 22701761
references.citation: Mol Cell Biol. 1993 Nov;13(11):6849-57
references.pmid: 8413278
references.citation: Nature. 1992 Mar 19;356(6366):215-21
references.pmid: 1552940
references.citation: Carcinogenesis. 2012 Feb;33(2):442-51
references.pmid: 22114072
references.citation: Cell. 2004 Dec 17;119(6):861-72
references.pmid: 15607981
references.citation: Genes Cancer. 2012 Jul;3(7-8):491-502
references.pmid: 23264849
references.citation: BMC Med. 2012 Mar 21;10 :28
references.pmid: 22436374
references.citation: Carcinogenesis. 1996 Jun;17(6):1187-98
references.pmid: 8681432