title: The KRAS promoter responds to Myc-associated zinc finger and poly(ADP-ribose) polymerase 1 proteins, which recognize a critical quadruplex-forming GA-element.
abstract: The murine KRAS promoter contains a G-rich nuclease hypersensitive element (GA-element) upstream of the transcription start site that is essential for transcription. Pulldown and chromatin immunoprecipitation assays demonstrate that this GA-element is bound by the Myc-associated zinc finger (MAZ) and poly(ADP-ribose) polymerase 1 (PARP-1) proteins. These proteins are crucial for transcription, because when they are knocked down by short hairpin RNA, transcription is down-regulated. This is also the case when the poly(ADP-ribosyl)ation activity of PARP-1 is inhibited by 3,4-dihydro-5-[4-(1-piperidinyl) butoxyl]-1(2H) isoquinolinone. We found that MAZ specifically binds to the duplex and quadruplex conformations of the GA-element, whereas PARP-1 shows specificity only for the G-quadruplex. On the basis of fluorescence resonance energy transfer melting and polymerase stop assays we saw that MAZ stabilizes the KRAS quadruplex. When the capacity of folding in the GA-element is abrogated by specific G --> T or G --> A point mutations, KRAS transcription is down-regulated. Conversely, guanidine-modified phthalocyanines, which specifically interact with and stabilize the KRAS G-quadruplex, push the promoter activity up to more than double. Collectively, our data support a transcription mechanism for murine KRAS that involves MAZ, PARP-1 and duplex-quadruplex conformational changes in the promoter GA-element.
authors.affiliation: Department of Biomedical Science and Technology, University of Udine, School of Medicine, Piazzale Kolbe 4, 33100 Udine, Italy.
authors.forename: Susanna
authors.initials: S
authors.lastname: Cogoi
authors.forename: Manikandan
authors.initials: M
authors.lastname: Paramasivam
authors.forename: Alexandro
authors.initials: A
authors.lastname: Membrino
authors.forename: Kazunari K
authors.initials: KK
authors.lastname: Yokoyama
authors.forename: Luigi E
authors.initials: LE
authors.lastname: Xodo
mesh_terms: D000818:Animals; D001483:Base Sequence; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D031541:Fluorescence Resonance Energy Transfer; D054856:G-Quadruplexes; D006147:Guanine / Q000378:metabolism*; D007211:Indoles / Q000737:chemistry; D054833:Isoindoles; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D041681:NIH 3T3 Cells; D009690:Nucleic Acid Conformation; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D055550:Protein Stability; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D004268:DNA-Binding Proteins; D007211:Indoles; D054833:Isoindoles; D008024:Ligands; D050505:Mutant Proteins; D014157:Transcription Factors; C076035:c-MYC-associated zinc finger protein; D006147:Guanine; D011065:Poly(ADP-ribose) Polymerases; C495971:Hras protein, mouse; D016283:Proto-Oncogene Proteins p21(ras); C013647:phthalocyanine
references.citation: J Cell Physiol. 2009 May;219(2):265-70
references.pmid: 19170077
references.citation: Science. 1999 Dec 17;286(5448):2339-42
references.pmid: 10600744
references.citation: BMC Genomics. 2007 Feb 07;8:41
references.pmid: 17286852
references.citation: Nat Rev Cancer. 2009 Dec;9(12):849-61
references.pmid: 19907434
references.citation: J Biol Chem. 2005 Apr 29;280(17):17076-83
references.pmid: 15737996
references.citation: Cancer Res. 2006 Jun 1;66(11):5744-56
references.pmid: 16740713
references.citation: J Biol Chem. 1999 Jan 29;274(5):3076-93
references.pmid: 9915847
references.citation: Nucleic Acids Res. 2008 Jan;36(1):144-56
references.pmid: 17999996
references.citation: Biochem Biophys Res Commun. 2001 Sep 7;286(5):1087-97
references.pmid: 11527412
references.citation: Nucleic Acids Res. 2009 Jan;37(1):172-83
references.pmid: 19033359
references.citation: J Med Chem. 2009 May 14;52(9):2863-74
references.pmid: 19385599
references.citation: Chem Commun (Camb). 2010 Jan 28;46(4):625-7
references.pmid: 20062883
references.citation: Nucleic Acids Res. 2005 May 24;33(9):2901-7
references.pmid: 15914666
references.citation: Mol Pharmacol. 2003 Nov;64(5):1180-8
references.pmid: 14573768
references.citation: Science. 2009 Aug 7;325(5941):764-7
references.pmid: 19661435
references.citation: Nat Struct Mol Biol. 2008 Feb;15(2):146-54
references.pmid: 18193062
references.citation: Biochemistry. 2000 Mar 28;39(12):3206-15
references.pmid: 10727212
references.citation: J Biol Chem. 2001 Jun 8;276(23):19897-904
references.pmid: 11259406
references.citation: Mol Cell Biol. 2007 Nov;27(21):7475-85
references.pmid: 17785446
references.citation: J Biol Chem. 1998 Aug 7;273(32):20603-14
references.pmid: 9685418
references.citation: Biochem Biophys Res Commun. 2009 Mar 13;380(3):431-6
references.pmid: 19284986
references.citation: Cell. 2003 Jun 13;113(6):677-83
references.pmid: 12809599
references.citation: Biochemistry. 1999 Dec 7;38(49):16067-75
references.pmid: 10587429
references.citation: Mol Cell Biol. 2008 Oct;28(20):6521-35
references.pmid: 18710939
references.citation: Chem Commun (Camb). 2009 Apr 21;(15):1970-1
references.pmid: 19333460
references.citation: Nucleic Acids Res. 1991 Dec 11;19(23):6527-32
references.pmid: 1754390
references.citation: Nature. 1988 Jul 28;334(6180):364-6
references.pmid: 3393228
references.citation: Nat Struct Mol Biol. 2006 Dec;13(12):1055-9
references.pmid: 17146462
references.citation: Proc Natl Acad Sci U S A. 1990 Apr;87(7):2705-9
references.pmid: 2181446
references.citation: Nucleic Acids Res. 2007;35(22):7698-713
references.pmid: 17984069
references.citation: J Mol Endocrinol. 2004 Feb;32(1):99-113
references.pmid: 14765995
references.citation: Proc Natl Acad Sci U S A. 2000 Nov 7;97(23):12508-12
references.pmid: 11070077
references.citation: Nucleic Acids Res. 2006;34(14):3887-96
references.pmid: 16914419
references.citation: Proc Natl Acad Sci U S A. 2002 Sep 3;99(18):11593-8
references.pmid: 12195017
references.citation: Mol Cell Biol. 1996 May;16(5):2350-60
references.pmid: 8628302
references.citation: Biochem Biophys Res Commun. 1996 Sep 24;226(3):801-9
references.pmid: 8831693
references.citation: Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7452-6
references.pmid: 1502157
references.citation: Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5830-5
references.pmid: 8650178
references.citation: J Am Chem Soc. 2005 Aug 3;127(30):10584-9
references.pmid: 16045346
references.citation: Nucleic Acids Res. 2008 Apr;36(6):1755-69
references.pmid: 18252774
references.citation: J Biol Chem. 1996 Feb 23;271(8):4417-30
references.pmid: 8626793
references.citation: Mol Cancer Ther. 2008 Apr;7(4):880-9
references.pmid: 18413801
references.citation: J Mol Biol. 2005 May 6;348(3):609-15
references.pmid: 15826658
references.citation: Biophys J. 2007 Mar 15;92(6):2007-15
references.pmid: 17172304
references.citation: Cell Mol Life Sci. 2005 Apr;62(7-8):739-50
references.pmid: 15868399
references.citation: Nucleic Acids Res. 2008 Jun;36(11):3765-80
references.pmid: 18490377
references.citation: Science. 2008 Feb 8;319(5864):819-21
references.pmid: 18258916
references.citation: Cancer Res. 2007 Nov 1;67(21):10491-500
references.pmid: 17974993
references.citation: J Virol. 1997 Dec;71(12):9600-7
references.pmid: 9371624
references.citation: Nucleic Acids Res. 2010 Mar;38(5):1623-35
references.pmid: 20008101
references.citation: Biochem Biophys Res Commun. 1999 Aug 19;262(1):198-205
references.pmid: 10448092
references.citation: Trends Biochem Sci. 1994 Apr;19(4):172-6
references.pmid: 8016868
references.citation: J Biol Chem. 1999 Jul 2;274(27):19498-506
references.pmid: 10383467
references.citation: Cytometry A. 2005 Nov;68(1):21-7
references.pmid: 16200639
references.citation: Nucleic Acids Res. 2007;35(2):406-13
references.pmid: 17169996
references.citation: Nucleic Acids Res. 2006 May 10;34(9):2536-49
references.pmid: 16687659
references.citation: J Am Chem Soc. 2003 Apr 2;125(13):3763-7
references.pmid: 12656607
references.citation: ACS Chem Biol. 2008 Apr 18;3(4):214-9
references.pmid: 18338862