Identification of the Adenovirus E4orf4 Protein Binding Site on the B55α and Cdc55 Regulatory Subunits of PP2A: Implications for PP2A Function, Tumor Cell Killing and Viral Replication
Adenovirus E4orf4 protein induces the death of human cancer cells and Saccharomyces cerevisiae. Binding of E4orf4 to the B/B55/Cdc55 regulatory subunit of protein phosphatase 2A (PP2A) is required, and such binding inhibits PP2AB55 activity leading to dose-dependent cell death. We found that E4orf4 binds across the putative substrate binding groove predicted from the crystal structure of B55α such that the substrate p107 can no longer interact with PP2AB55α. We propose that E4orf4 inhibits PP2AB55 activity by preventing access of substrates and that at high E4orf4 levels this inhibition results in cell death through the failure to dephosphorylate substrates required for cell cycle progression. However, E4orf4 is expressed at much lower and less toxic levels during a normal adenovirus infection. We suggest that in this context E4orf4 largely serves to recruit novel substrates such as ASF/SF2/SRSF1 to PP2AB55 to enhance adenovirus replication. Thus E4orf4 toxicity probably represents an artifact of overexpression and does not reflect the evolutionary function of this viral product.
Vyšlo v časopise:
Identification of the Adenovirus E4orf4 Protein Binding Site on the B55α and Cdc55 Regulatory Subunits of PP2A: Implications for PP2A Function, Tumor Cell Killing and Viral Replication. PLoS Pathog 9(11): e32767. doi:10.1371/journal.ppat.1003742
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1003742
Souhrn
Adenovirus E4orf4 protein induces the death of human cancer cells and Saccharomyces cerevisiae. Binding of E4orf4 to the B/B55/Cdc55 regulatory subunit of protein phosphatase 2A (PP2A) is required, and such binding inhibits PP2AB55 activity leading to dose-dependent cell death. We found that E4orf4 binds across the putative substrate binding groove predicted from the crystal structure of B55α such that the substrate p107 can no longer interact with PP2AB55α. We propose that E4orf4 inhibits PP2AB55 activity by preventing access of substrates and that at high E4orf4 levels this inhibition results in cell death through the failure to dephosphorylate substrates required for cell cycle progression. However, E4orf4 is expressed at much lower and less toxic levels during a normal adenovirus infection. We suggest that in this context E4orf4 largely serves to recruit novel substrates such as ASF/SF2/SRSF1 to PP2AB55 to enhance adenovirus replication. Thus E4orf4 toxicity probably represents an artifact of overexpression and does not reflect the evolutionary function of this viral product.
Zdroje
1. MarcellusRC, TeodoroJG, WuT, BroughDE, KetnerG, et al. (1996) Adenovirus type 5 early region 4 is responsible for E1A-induced p53-independent apoptosis. Journal Of Virology 70: 6207–6215.
2. LavoieJN, NguyenM, MarcellusRC, BrantonPE, ShoreGC (1998) E4orf4, a novel adenovirus death factor that induces p53-independent apoptosis by a pathway that is not inhibited by zVAD-fmk. Journal Of Cell Biology 140: 637–645.
3. MarcellusRC, LavoieJN, BoivinD, ShoreGC, KetnerG, et al. (1998) The early region 4 orf4 protein of human adenovirus type 5 induces p53-independent cell death by apoptosis. Journal Of Virology 72: 7144–7153.
4. ShtrichmanR, KleinbergerT (1998) Adenovirus type 5 E4 open reading frame 4 protein induces apoptosis in transformed cells. Journal Of Virology 72: 2975–2982.
5. ShtrichmanR, SharfR, BarrH, DobnerT, KleinbergerT (1999) Induction of apoptosis by adenovirus E4orf4 protein is specific to transformed cells and requires an interaction with protein phosphatase 2A. Proceedings Of The National Academy Of Sciences Of The United States Of America 96: 10080–10085.
6. LavoieJN, ChampagneC, GingrasMC, RobertA (2000) Adenovirus E4 open reading frame 4-induced apoptosis involves dysregulation of Src family kinases. Journal of Cell Biology 150: 1037–1056.
7. MarcellusRC, ChanH, PaquetteD, ThirlwellS, BoivinD, et al. (2000) Induction of p53-independent apoptosis by the adenovirus E4orf4 protein requires binding to the Balpha subunit of protein phosphatase 2A. J Virol 74: 7869–7877.
8. ShtrichmanR, SharfR, KleinbergerT (2000) Adenovirus E4orf4 protein interacts with both Balpha and B′ subunits of protein phosphatase 2A, but E4orf4-induced apoptosis is mediated only by the interaction with Balpha. Oncogene 19: 3757–3765.
9. KornitzerD, SharfR, KleinbergerT (2001) Adenovirus E4orf4 protein induces PP2A-dependent growth arrest in Saccharomyces cerevisiae and interacts with the anaphase-promoting complex/cyclosome. J Cell Biol 154: 331–344.
10. LivneA, ShtrichmanR, KleinbergerT (2001) Caspase activation by adenovirus e4orf4 protein is cell line specific and Is mediated by the death receptor pathway. Journal of Virology 75: 789–798.
11. RoopchandDE, LeeJM, ShahinianS, PaquetteD, BusseyH, et al. (2001) Toxicity of human adenovirus E4orf4 protein in Saccharomyces cerevisiae results from interactions with the Cdc55 regulatory B subunit of PP2A. Oncogene 20: 5279–5290.
12. LiS, BrignoleC, MarcellusR, ThirlwellS, BindaO, et al. (2009) The adenovirus E4orf4 protein induces G2/M arrest and cell death by blocking protein phosphatase 2A activity regulated by the B55 subunit. J Virol 83: 8340–8352.
13. LiS, SzymborskiA, MironMJ, MarcellusR, BindaO, et al. (2009) The adenovirus E4orf4 protein induces growth arrest and mitotic catastrophe in H1299 human lung carcinoma cells. Oncogene 28: 390–400.
14. MuiMZ, RoopchandDE, GentryMS, HallbergRL, VogelJ, et al. (2010) Adenovirus protein E4orf4 induces premature APCCdc20 activation in Saccharomyces cerevisiae by a protein phosphatase 2A-dependent mechanism. J Virol 84: 4798–4809.
15. LiY, WeiH, HsiehTC, PallasDC (2008) Cdc55p-mediated E4orf4 growth inhibition in Saccharomyces cerevisiae is mediated only in part via the catalytic subunit of protein phosphatase 2A. J Virol 82: 3612–3623.
16. RobertA, MironMJ, ChampagneC, GingrasMC, BrantonPE, et al. (2002) Distinct cell death pathways triggered by the adenovirus early region 4 ORF 4 protein. J Cell Biol 158: 519–528.
17. PechkovskyA, LahavM, BitmanE, SalzbergA, KleinbergerT (2013) E4orf4 induces PP2A- and Src-dependent cell death in Drosophila melanogaster and at the same time inhibits classic apoptosis pathways. Proceedings Of The National Academy Of Sciences Of The United States Of America 110: E1724–1733.
18. CabonL, SriskandarajahN, MuiMZ, TeodoroJG, BlanchetteP, et al. (2013) Adenovirus E4orf4 Protein-Induced Death of p53−/− H1299 Human Cancer Cells Follows a G1 Arrest of Both Tetraploid and Diploid Cells Due to a Failure to Initiate DNA Synthesis. Journal Of Virology 01242–13.
19. BrestovitskyA, SharfR, MittelmanK, KleinbergerT (2011) The adenovirus E4orf4 protein targets PP2A to the ACF chromatin-remodeling factor and induces cell death through regulation of SNF2h-containing complexes. Nucleic acids research 39: 6414–6427.
20. KleinbergerT, ShenkT (1993) Adenovirus E4orf4 protein binds to protein phosphatase 2A, and the complex down regulates E1A-enhanced junB transcription. Journal Of Virology 67: 7556–7560.
21. KorenR, RainisL, KleinbergerT (2004) The scaffolding A/Tpd3 subunit and high phosphatase activity are dispensable for Cdc55 function in the Saccharomyces cerevisiae spindle checkpoint and in cytokinesis. J Biol Chem 279: 48598–48606.
22. AfifiR, SharfR, ShtrichmanR, KleinbergerT (2001) Selection of apoptosis-deficient adenovirus E4orf4 mutants in Saccharomyces cerevisiae. Journal of Virology 75: 4444–4447.
23. HorowitzB, SharfR, Avital-ShachamM, PechkovskyA, KleinbergerT (2013) Structure- and modeling-based identification of the adenovirus E4orf4 binding site in the protein phosphatase 2A (PP2A) B55alpha subunit. journal of biological chemistry 288: 13718–13727.
24. ZhangZ, MuiMZ, ChanF, RoopchandDE, MarcellusRC, et al. (2011) Genetic analysis of B55alpha/Cdc55 protein phosphatase 2A subunits: association with the adenovirus E4orf4 protein. J Virol 85: 286–295.
25. MumbyMC, WalterG (1993) Protein serine/threonine phosphatases: structure, regulation, and functions in cell growth. physiological reviews 73: 673–699.
26. StarkMJR (1996) Yeast protein serine threonine phosphatases: multiple roles and diverse regulation. Yeast 12: 1647–1675.
27. GoldbergY (1999) Protein Phosphatase 2A: Who Shall Regulate the Regulator. Biochemical Pharmacology 57: 321–328.
28. LechwardK, ZolnierowiczS, HemmingsBA (1999) Eukaryotic translation termination factor 1 associates with protein phosphatase 2A and targets it to ribosomes. biochemistry (mosc) 64: 1373–1381.
29. JanssensV, GorisJ (2001) Protein phosphatase 2A: a highly regulated family of serine/threonine phosphatases implicated in cell growth and signalling. Biochemical Journal 353: 417–439.
30. KurimchakA, GranaX (2012) PP2A holoenzymes negatively and positively regulate cell cycle progression by dephosphorylating pocket proteins and multiple CDK substrates. Gene 499: 1–7.
31. SablinaAA, HectorM, ColpaertN, HahnWC (2010) Identification of PP2A complexes and pathways involved in cell transformation. Cancer Res 70: 10474–10484.
32. WestermarckJ, HahnWC (2008) Multiple pathways regulated by the tumor suppressor PP2A in transformation. Trends Mol Med 14: 152–160.
33. FerrignoP, LanganTA, CohenP (1993) Protein phosphatase 2A1 is the major enzyme in vertebrate cell extracts that dephosphorylates several physiological substrates for cyclin-dependent protein kinases. molecular biology of the cell 4: 669–677.
34. KamibayashiC, EstesR, LickteigRL, YangSI, CraftC, et al. (1994) Comparison of heterotrimeric protein phosphatase 2A containing different B subunits. journal of biological chemistry 269: 20139–20148.
35. JanssensV, LonginS, GorisJ (2008) PP2A holoenzyme assembly: in cauda venenum (the sting is in the tail). Trends Biochem Sci 33: 113–121.
36. SentsW, IvanovaE, LambrechtC, HaesenD, JanssensV (2013) The biogenesis of active protein phosphatase 2A holoenzymes: a tightly regulated process creating phosphatase specificity. FEBS J 280: 644–661.
37. VirshupDM, ShenolikarS (2009) From promiscuity to precision: protein phosphatases get a makeover. Mol Cell 33: 537–545.
38. SneddonAA, CohenPT, StarkMJ (1990) Saccharomyces cerevisiae protein phosphatase 2A performs an essential cellular function and is encoded by two genes. embo journal 9: 4339–4346.
39. RonneH, CarlbergM, HuGZ, NehlinJO (1991) Protein phosphatase 2A in Saccharomyces cerevisiae: effects on cell growth and bud morphogenesis. Molecular And Cellular Biology 11: 4876–4884.
40. van ZylW, HuangW, SneddonAA, StarkM, CamierS, et al. (1992) Inactivation of the protein phosphatase 2A regulatory subunit A results in morphological and transcriptional defects in Saccharomyces cerevisiae. Molecular And Cellular Biology 12: 4946–4959.
41. ShuY, YangH, HallbergE, HallbergR (1997) Molecular genetic analysis of Rts1p, a B′ regulatory subunit of Saccharomyces cerevisiae protein phosphatase 2A. Molecular And Cellular Biology 17: 3242–3253.
42. HealyAM, ZolnierowiczS, StapletonAE, GoeblM, DePaoli-RoachAA, et al. (1991) CDC55, a Saccharomyces cerevisiae gene involved in cellular morphogenesis: identification, characterization, and homology to the B subunit of mammalian type 2A protein phosphatase. Molecular And Cellular Biology 11: 5767–5780.
43. ArroyoJD, HahnWC (2005) Involvement of PP2A in viral and cellular transformation. Oncogene 24: 7746–7755.
44. PallasDC, CheringtonV, MorganW, DeAndaJ, KaplanD, et al. (1988) Cellular proteins that associate with the middle and small T antigens of polyomavirus. Journal Of Virology 62: 3934–3940.
45. PallasDC, ShahrikLK, MartinBL, JaspersS, MillerTB, et al. (1990) Polyoma small and middle T antigens and SV40 small t antigen form stable complexes with protein phosphatase 2A. Cell 60: 167–176.
46. WalterG, RuedigerR, SlaughterC, MumbyM (1990) Association of protein phosphatase 2A with polyoma virus medium tumor antigen. Proceedings Of The National Academy Of Sciences Of The United States Of America 87: 2521–2525.
47. YangSI, LickteigRL, EstesR, RundellK, WalterG, et al. (1991) Control of protein phosphatase 2A by simian virus 40 small-t antigen. molecular and cellular biology 11: 1988–1995.
48. RuedigerR, RoeckelD, FaitJ, BergqvistA, MagnussonG, et al. (1992) Identification of binding sites on the regulatory A subunit of protein phosphatase 2A for the catalytic C subunit and for tumor antigens of simian virus 40 and polyomavirus. molecular and cellular biology 12: 4872–4882.
49. MironMJ, GallouziIE, LavoieJN, BrantonPE (2004) Nuclear localization of the adenovirus E4orf4 protein is mediated through an arginine-rich motif and correlates with cell death. Oncogene 23: 7458–7468.
50. XuY, ChenY, ZhangP, JeffreyPD, ShiY (2008) Structure of a protein phosphatase 2A holoenzyme: insights into B55-mediated Tau dephosphorylation. Mol Cell 31: 873–885.
51. XingY, XuY, ChenY, JeffreyPD, ChaoY, et al. (2006) Structure of protein phosphatase 2A core enzyme bound to tumor-inducing toxins. Cell 127: 341–353.
52. O'SheaC, KlupschK, ChoiS, BagusB, SoriaC, et al. (2005) Adenoviral proteins mimic nutrient/growth signals to activate the mTOR pathway for viral replication. The EMBO journal 24: 1211–1221.
53. KleinbergerT (2004) Induction of transformed cell-specific apoptosis by the adenovirus E4orf4 protein. Prog Mol Subcell Biol 36: 245–267.
54. JayadevaG, KurimchakA, GarrigaJ, SotilloE, DavisAJ, et al. (2010) B55alpha PP2A holoenzymes modulate the phosphorylation status of the retinoblastoma-related protein p107 and its activation. J Biol Chem 285: 29863–29873.
55. KurimchakA, HainesDS, GarrigaJ, WuS, De LucaF, et al. (2013) Activation of p107 by FGF, which is essential for chondrocyte cell cycle exit, is mediated by the PP2A/B55alpha holoenzyme. molecular and cellular biology doi:10.1128/MCB.00082-13
56. KolupaevaV, DaempflingL, BasilicoC (2013) The B55 alpha regulatory subunit of Protein Phosphatase 2A mediates FGF-induced p107 dephosphorylation and growth arrest in chondrocytes. molecular and cellular biology doi:10.1128/MCB.01730-12
57. MironMJ, BlanchetteP, GroitlP, DallaireF, TeodoroJG, et al. (2009) Localization and importance of the adenovirus E4orf4 protein during lytic infection. Journal Of Virology 83: 1689–1699.
58. KanopkaA, MühlemannO, Petersen-MahrtS, EstmerC, OhrmalmC, et al. (1998) Regulation of adenovirus alternative RNA splicing by dephosphorylation of SR proteins. Nature 393: 185–187.
59. Estmer NilssonC, Petersen-MahrtS, DurotC, ShtrichmanR, KrainerAR, et al. (2001) The adenovirus E4-ORF4 splicing enhancer protein interacts with a subset of phosphorylated SR proteins. The EMBO journal 20: 864–871.
60. ChenDC, YangBC, KuoTT (1992) One-step transformation of yeast in stationary phase. Current Genetics 21: 83–84.
61. SaliA, BlundellTL (1993) Comparative protein modelling by satisfaction of spatial restraints. J Mol Biol 234: 779–815.
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Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
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