Beta-HPV 5 and 8 E6 Promote p300 Degradation by Blocking AKT/p300 Association


The E6 oncoprotein from high-risk genus alpha human papillomaviruses (α-HPVs), such as HPV 16, has been well characterized with respect to the host-cell proteins it interacts with and corresponding signaling pathways that are disrupted due to these interactions. Less is known regarding the interacting partners of E6 from the genus beta papillomaviruses (β-HPVs); however, it is generally thought that β-HPV E6 proteins do not interact with many of the proteins known to bind to α-HPV E6. Here we identify p300 as a protein that interacts directly with E6 from both α- and β-HPV types. Importantly, this association appears much stronger with β-HPV types 5 and 8-E6 than with α-HPV type 16-E6 or β-HPV type 38-E6. We demonstrate that the enhanced association between 5/8-E6 and p300 leads to p300 degradation in a proteasomal-dependent but E6AP-independent manner. Rather, 5/8-E6 inhibit the association of AKT with p300, an event necessary to ensure p300 stability within the cell. Finally, we demonstrate that the decreased p300 protein levels concomitantly affect downstream signaling events, such as the expression of differentiation markers K1, K10 and Involucrin. Together, these results demonstrate a unique way in which β-HPV E6 proteins are able to affect host-cell signaling in a manner distinct from that of the α-HPVs.


Vyšlo v časopise: Beta-HPV 5 and 8 E6 Promote p300 Degradation by Blocking AKT/p300 Association. PLoS Pathog 7(8): e32767. doi:10.1371/journal.ppat.1002211
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002211

Souhrn

The E6 oncoprotein from high-risk genus alpha human papillomaviruses (α-HPVs), such as HPV 16, has been well characterized with respect to the host-cell proteins it interacts with and corresponding signaling pathways that are disrupted due to these interactions. Less is known regarding the interacting partners of E6 from the genus beta papillomaviruses (β-HPVs); however, it is generally thought that β-HPV E6 proteins do not interact with many of the proteins known to bind to α-HPV E6. Here we identify p300 as a protein that interacts directly with E6 from both α- and β-HPV types. Importantly, this association appears much stronger with β-HPV types 5 and 8-E6 than with α-HPV type 16-E6 or β-HPV type 38-E6. We demonstrate that the enhanced association between 5/8-E6 and p300 leads to p300 degradation in a proteasomal-dependent but E6AP-independent manner. Rather, 5/8-E6 inhibit the association of AKT with p300, an event necessary to ensure p300 stability within the cell. Finally, we demonstrate that the decreased p300 protein levels concomitantly affect downstream signaling events, such as the expression of differentiation markers K1, K10 and Involucrin. Together, these results demonstrate a unique way in which β-HPV E6 proteins are able to affect host-cell signaling in a manner distinct from that of the α-HPVs.


Zdroje

1. BernardHUBurkRDChenZvan DoorslaerKHausenH 2010 Classification of papillomaviruses (PVs) based on 189 PV types and proposal of taxonomic amendments. Virology 401 70 79

2. zur HausenH 1999 Papillomaviruses in human cancers. Proc Assoc Am Physicians 111 581 587

3. CoglianoVBaanRStraifKGrosseYSecretanB 2005 Carcinogenicity of human papillomaviruses. Lancet Oncol 6 204

4. Bouwes BavinckJNFeltkampMStruijkLter ScheggetJ 2001 Human papillomavirus infection and skin cancer risk in organ transplant recipients. J Investig Dermatol Symp Proc 6 207 211

5. OrthGJablonskaSJarzabek-ChorzelskaMObalekSRzesaG 1979 Characteristics of the lesions and risk of malignant conversion associated with the type of human papillomavirus involved in epidermodysplasia verruciformis. Cancer Res 39 1074 1082

6. LutznerMABlanchet-BardonCOrthG 1984 Clinical observations, virologic studies, and treatment trials in patients with epidermodysplasia verruciformis, a disease induced by specific human papillomaviruses. J Invest Dermatol 83 18s 25s

7. PfisterH 1992 Human papillomaviruses and skin cancer. Semin Cancer Biol 3 263 271

8. HowieHLKatzenellenbogenRAGallowayDA 2009 Papillomavirus E6 proteins. Virology 384 324 334

9. McLaughlin-DrubinMEMungerK 2009 The human papillomavirus E7 oncoprotein. Virology 384 335 344

10. HuibregtseJMScheffnerMHowleyPM 1991 A cellular protein mediates association of p53 with the E6 oncoprotein of human papillomavirus types 16 or 18. Embo J 10 4129 4135

11. ScheffnerMHuibregtseJMVierstraRDHowleyPM 1993 The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53. Cell 75 495 505

12. ChenJJHongYRustamzadehEBalejaJDAndrophyEJ 1998 Identification of an alpha helical motif sufficient for association with papillomavirus E6. J Biol Chem 273 13537 13544

13. KuballaPMatentzogluKScheffnerM 2007 The role of the ubiquitin ligase E6-AP in human papillomavirus E6-mediated degradation of PDZ domain-containing proteins. J Biol Chem 282 65 71

14. ThomasMBanksL 1999 Human papillomavirus (HPV) E6 interactions with Bak are conserved amongst E6 proteins from high and low risk HPV types. J Gen Virol 80 Pt 6 1513 1517

15. ThomasMBanksL 1998 Inhibition of Bak-induced apoptosis by HPV-18 E6. Oncogene 17 2943 2954

16. KiyonoTFosterSAKoopJIMcDougallJKGallowayDA 1998 Both Rb/p16INK4a inactivation and telomerase activity are required to immortalize human epithelial cells. Nature 396 84 88

17. KlingelhutzAJFosterSAMcDougallJK 1996 Telomerase activation by the E6 gene product of human papillomavirus type 16. Nature 380 79 82

18. VeldmanTLiuXYuanHSchlegelR 2003 Human papillomavirus E6 and Myc proteins associate in vivo and bind to and cooperatively activate the telomerase reverse transcriptase promoter. Proc Natl Acad Sci U S A 100 8211 8216

19. ScheffnerMWernessBAHuibregtseJMLevineAJHowleyPM 1990 The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53. Cell 63 1129 1136

20. KiyonoTHiraiwaAFujitaMHayashiYAkiyamaT 1997 Binding of high-risk human papillomavirus E6 oncoproteins to the human homologue of the Drosophila discs large tumor suppressor protein. Proc Natl Acad Sci U S A 94 11612 11616

21. LeeSSWeissRSJavierRT 1997 Binding of human virus oncoproteins to hDlg/SAP97, a mammalian homolog of the Drosophila discs large tumor suppressor protein. Proc Natl Acad Sci U S A 94 6670 6675

22. NakagawaSHuibregtseJM 2000 Human scribble (Vartul) is targeted for ubiquitin-mediated degradation by the high-risk papillomavirus E6 proteins and the E6AP ubiquitin-protein ligase. Mol Cell Biol 20 8244 8253

23. GlaunsingerBALeeSSThomasMBanksLJavierR 2000 Interactions of the PDZ-protein MAGI-1 with adenovirus E4-ORF1 and high-risk papillomavirus E6 oncoproteins. Oncogene 19 5270 5280

24. ThomasMGlaunsingerBPimDJavierRBanksL 2001 HPV E6 and MAGUK protein interactions: determination of the molecular basis for specific protein recognition and degradation. Oncogene 20 5431 5439

25. LeeSSGlaunsingerBMantovaniFBanksLJavierRT 2000 Multi-PDZ domain protein MUPP1 is a cellular target for both adenovirus E4-ORF1 and high-risk papillomavirus type 18 E6 oncoproteins. J Virol 74 9680 9693

26. JingMBohlJBrimerNKinterMVande PolSB 2007 Degradation of tyrosine phosphatase PTPN3 (PTPH1) by association with oncogenic human papillomavirus E6 proteins. J Virol 81 2231 2239

27. SpanosWCHooverAHarrisGFWuSStrandGL 2008 The PDZ binding motif of human papillomavirus type 16 E6 induces PTPN13 loss, which allows anchorage-independent growth and synergizes with ras for invasive growth. J Virol 82 2493 2500

28. ThomasMDasguptaJZhangYChenXBanksL 2008 Analysis of specificity determinants in the interactions of different HPV E6 proteins with their PDZ domain-containing substrates. Virology 376 371 378

29. UnderbrinkMPHowieHLBedardKMKoopJIGallowayDA 2008 E6 proteins from multiple human betapapillomavirus types degrade Bak and protect keratinocytes from apoptosis after UVB irradiation. J Virol 82 10408 10417

30. BedardKMUnderbrinkMPHowieHLGallowayDA 2008 The E6 oncoproteins from human betapapillomaviruses differentially activate telomerase through an E6AP-dependent mechanism and prolong the lifespan of primary keratinocytes. J Virol 82 3894 3902

31. AccardiRDongWSmetACuiRHautefeuilleA 2006 Skin human papillomavirus type 38 alters p53 functions by accumulation of deltaNp73. EMBO Rep 7 334 340

32. DongWArpinCAccardiRGissmannLSyllaBS 2008 Loss of p53 or p73 in human papillomavirus type 38 E6 and E7 transgenic mice partially restores the UV-activated cell cycle checkpoints. Oncogene 27 2923 2928

33. Muller-SchiffmannABeckmannJStegerG 2006 The E6 protein of the cutaneous human papillomavirus type 8 can stimulate the viral early and late promoters by distinct mechanisms. J Virol 80 8718 8728

34. ZimmermannHDegenkolbeRBernardHUO'ConnorMJ 1999 The human papillomavirus type 16 E6 oncoprotein can down-regulate p53 activity by targeting the transcriptional coactivator CBP/p300. J Virol 73 6209 6219

35. ZimmermannHKohCHDegenkolbeRO'ConnorMJMullerA 2000 Interaction with CBP/p300 enables the bovine papillomavirus type 1 E6 oncoprotein to downregulate CBP/p300-mediated transactivation by p53. J Gen Virol 81 2617 2623

36. PatelDHuangSMBagliaLAMcCanceDJ 1999 The E6 protein of human papillomavirus type 16 binds to and inhibits co-activation by CBP and p300. Embo J 18 5061 5072

37. MuenchPProbstSSchuetzJLeiprechtNBuschM 2010 Cutaneous Papillomavirus E6 Proteins Must Interact with p300 and Block p53-Mediated Apoptosis for Cellular Immortalization and Tumorigenesis. Cancer Res 70 6913 6924

38. ThomasMCChiangCM 2005 E6 oncoprotein represses p53-dependent gene activation via inhibition of protein acetylation independently of inducing p53 degradation. Mol Cell 17 251 264

39. GilesRHPetersDJBreuningMH 1998 Conjunction dysfunction: CBP/p300 in human disease. Trends Genet 14 178 183

40. GoodmanRHSmolikS 2000 CBP/p300 in cell growth, transformation, and development. Genes Dev 14 1553 1577

41. KaoWHBeaudenonSLTalisALHuibregtseJMHowleyPM 2000 Human papillomavirus type 16 E6 induces self-ubiquitination of the E6AP ubiquitin-protein ligase. J Virol 74 6408 6417

42. ChenJHalappanavarSSSt-GermainJRTsangBKLiQ 2004 Role of Akt/protein kinase B in the activity of transcriptional coactivator p300. Cell Mol Life Sci 61 1675 1683

43. HuangWCChenCC 2005 Akt phosphorylation of p300 at Ser-1834 is essential for its histone acetyltransferase and transcriptional activity. Mol Cell Biol 25 6592 6602

44. GuoSCichySBHeXYangQRaglandM 2001 Insulin suppresses transactivation by CAAT/enhancer-binding proteins beta (C/EBPbeta). Signaling to p300/CREB-binding protein by protein kinase B disrupts interaction with the major activation domain of C/EBPbeta. J Biol Chem 276 8516 8523

45. GuptaSStuffreinSPlattnerRTencatiMGrayC 2001 Role of phosphoinositide 3-kinase in the aggressive tumor growth of HT1080 human fibrosarcoma cells. Mol Cell Biol 21 5846 5856

46. WenHCHuangWCAliAWoodgettJRLinWW 2003 Negative regulation of phosphatidylinositol 3-kinase and Akt signalling pathway by PKC. Cell Signal 15 37 45

47. VlahosCJMatterWFHuiKYBrownRF 1994 A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J Biol Chem 269 5241 5248

48. WongPPPickardAMcCanceDJ 2010 p300 alters keratinocyte cell growth and differentiation through regulation of p21(Waf1/CIP1). PLoS One 5 e8369

49. CrishJFEckertRL 2008 Synergistic activation of human involucrin gene expression by Fra-1 and p300—evidence for the presence of a multiprotein complex. J Invest Dermatol 128 530 541

50. SinitsynaNNReznikovaTVQinQSongHPhillipsMA 2010 Arsenite suppression of involucrin transcription through AP1 promoter sites in cultured human keratinocytes. Toxicol Appl Pharmacol 243 275 282

51. IyerNGOzdagHCaldasC 2004 p300/CBP and cancer. Oncogene 23 4225 4231

52. KalkhovenE 2004 CBP and p300: HATs for different occasions. Biochem Pharmacol 68 1145 1155

53. GaytherSABatleySJLingerLBannisterAThorpeK 2000 Mutations truncating the EP300 acetylase in human cancers. Nat Genet 24 300 303

54. BryanEJJokubaitisVJChamberlainNLBaxterSWDawsonE 2002 Mutation analysis of EP300 in colon, breast and ovarian carcinomas. Int J Cancer 102 137 141

55. GarbatiMRAlcoGGilmoreTD 2009 Histone acetyltransferase p300 is a coactivator for transcription factor REL and is C-terminally truncated in the human diffuse large B-cell lymphoma cell line RC-K8. Cancer Lett 291 237 245

56. MuraokaMKonishiMKikuchi-YanoshitaRTanakaKShitaraN 1996 p300 gene alterations in colorectal and gastric carcinomas. Oncogene 12 1565 1569

57. SuganumaTKawabataMOhshimaTIkedaMA 2002 Growth suppression of human carcinoma cells by reintroduction of the p300 coactivator. Proc Natl Acad Sci U S A 99 13073 13078

58. RebelVIKungALTannerEAYangHBronsonRT 2002 Distinct roles for CREB-binding protein and p300 in hematopoietic stem cell self-renewal. Proc Natl Acad Sci U S A 99 14789 14794

59. AranyZNewsomeDOldreadELivingstonDMEcknerR 1995 A family of transcriptional adaptor proteins targeted by the E1A oncoprotein. Nature 374 81 84

60. ChakravartiDOgryzkoVKaoHYNashAChenH 1999 A viral mechanism for inhibition of p300 and PCAF acetyltransferase activity. Cell 96 393 403

61. ColginMANyborgJK 1998 The human T-cell leukemia virus type 1 oncoprotein Tax inhibits the transcriptional activity of c-Myb through competition for the CREB binding protein. J Virol 72 9396 9399

62. HamamoriYSartorelliVOgryzkoVPuriPLWuHY 1999 Regulation of histone acetyltransferases p300 and PCAF by the bHLH protein twist and adenoviral oncoprotein E1A. Cell 96 405 413

63. KwokRPLauranceMELundbladJRGoldmanPSShihH 1996 Control of cAMP-regulated enhancers by the viral transactivator Tax through CREB and the co-activator CBP. Nature 380 642 646

64. GranjaAGNogalMLHurtadoCDel AguilaCCarrascosaAL 2006 The viral protein A238L inhibits TNF-alpha expression through a CBP/p300 transcriptional coactivators pathway. J Immunol 176 451 462

65. GranjaAGSanchezEGSabinaPFresnoMRevillaY 2009 African swine fever virus blocks the host cell antiviral inflammatory response through a direct inhibition of PKC-theta-mediated p300 transactivation. J Virol 83 969 980

66. KasperLHLerachSWangJWuSJeevanT 2010 CBP/p300 double null cells reveal effect of coactivator level and diversity on CREB transactivation. Embo J 29 3660 3672

67. KrubasikDIyerNGEnglishWRAhmedAAViasM 2006 Absence of p300 induces cellular phenotypic changes characteristic of epithelial to mesenchyme transition. Br J Cancer 94 1326 1332

68. YuanZMHuangYIshikoTNakadaSUtsugisawaT 1999 Function for p300 and not CBP in the apoptotic response to DNA damage. Oncogene 18 5714 5717

69. JamesMALeeJHKlingelhutzAJ 2006 HPV16-E6 associated hTERT promoter acetylation is E6AP dependent, increased in later passage cells and enhanced by loss of p300. Int J Cancer 119 1878 1885

70. SantosMParamioJMBravoARamirezAJorcanoJL 2002 The expression of keratin k10 in the basal layer of the epidermis inhibits cell proliferation and prevents skin tumorigenesis. J Biol Chem 277 19122 19130

71. ParamioJMCasanovaMLSegrellesCMittnachtSLaneEB 1999 Modulation of cell proliferation by cytokeratins K10 and K16. Mol Cell Biol 19 3086 3094

72. ParamioJMSegrellesCRuizSJorcanoJL 2001 Inhibition of protein kinase B (PKB) and PKCzeta mediates keratin K10-induced cell cycle arrest. Mol Cell Biol 21 7449 7459

73. AkgulBGhaliLDaviesDPfisterHLeighIM 2007 HPV8 early genes modulate differentiation and cell cycle of primary human adult keratinocytes. Exp Dermatol 16 590 599

74. BarcelosACSottoMN 2009 Comparative analysis of the expression of cytokeratins (1, 10, 14, 16, 4), involucrin, filaggrin and e-cadherin in plane warts and epidermodysplasia verruciformis plane wart-type lesions. J Cutan Pathol 36 647 654

75. SakaguchiKHerreraJESaitoSMikiTBustinM 1998 DNA damage activates p53 through a phosphorylation-acetylation cascade. Genes Dev 12 2831 2841

76. DornanDShimizuHPerkinsNDHuppTR 2003 DNA-dependent acetylation of p53 by the transcription coactivator p300. J Biol Chem 278 13431 13441

77. YuanZMHuangYIshikoTNakadaSUtsugisawaT 1999 Role for p300 in stabilization of p53 in the response to DNA damage. J Biol Chem 274 1883 1886

78. BarlevNALiuLChehabNHMansfieldKHarrisKG 2001 Acetylation of p53 activates transcription through recruitment of coactivators/histone acetyltransferases. Mol Cell 8 1243 1254

79. EspinosaJMEmersonBM 2001 Transcriptional regulation by p53 through intrinsic DNA/chromatin binding and site-directed cofactor recruitment. Mol Cell 8 57 69

80. CazzaliniOPeruccaPSavioMNecchiDBianchiL 2008 Interaction of p21(CDKN1A) with PCNA regulates the histone acetyltransferase activity of p300 in nucleotide excision repair. Nucleic Acids Res 36 1713 1722

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2011 Číslo 8
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Eozinofilní granulomatóza s polyangiitidou
nový kurz
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa