PUL21a-Cyclin A2 Interaction is Required to Protect Human Cytomegalovirus-Infected Cells from the Deleterious Consequences of Mitotic Entry

English version České info

Cyclin A2 is a key regulator of the cell division cycle. Interactors of Cyclin A2 typically contain short sequence elements (RXL/Cy motifs) that bind with high affinity to a hydrophobic patch in the Cyclin A2 protein. Two types of RXL/Cy-containing factors are known: i) cyclin-dependent kinase (CDK) substrates, which are processed by the CDK subunit that complexes to Cyclin A2, and ii) CDK inhibitors, which stably associate to Cyclin A2-CDK due to the lack of CDK phosphorylation sites. Human cytomegalovirus (HCMV) has evolved a novel type of RXL/Cy-containing protein. Its UL21a gene product, a small and highly unstable protein, binds to Cyclin A2 via an RXL/Cy motif in its N-terminus, leading to efficient degradation of Cyclin A2 by the proteasome. Here, we show that this mechanism is not only essential for viral inhibition of cellular DNA synthesis, but also to prevent entry of infected cells into mitosis. Unscheduled mitotic entry is followed by aberrant spindle formation, metaphase arrest, precocious separation of sister chromatids, chromosomal fragmentation and cell death. Viral DNA replication and expression of the essential viral IE2 protein are abrogated in mitosis. Thus, pUL21a-Cyclin A2 interaction protects HCMV from a collapse of viral and cellular functions in mitosis.

Vyšlo v časopise: PUL21a-Cyclin A2 Interaction is Required to Protect Human Cytomegalovirus-Infected Cells from the Deleterious Consequences of Mitotic Entry. PLoS Pathog 10(11): e32767. doi:10.1371/journal.ppat.1004514
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004514

Souhrn

Zdroje

1. BrittW (2008) Manifestations of human cytomegalovirus infection: proposed mechanisms of acute and chronic disease. Curr Top Microbiol Immunol 325: 417–470.

2. JohnsenJI, BaryawnoN, Soderberg-NauclerC (2011) Is human cytomegalovirus a target in cancer therapy? Oncotarget 2: 1329–1338.

3. SchreiberA, HarterG, SchubertA, BunjesD, MertensT, et al. (2009) Antiviral treatment of cytomegalovirus infection and resistant strains. Expert Opin Pharmacother 10: 191–209.

4. YuY, ClippingerAJ, AlwineJC (2011) Viral effects on metabolism: changes in glucose and glutamine utilization during human cytomegalovirus infection. Trends Microbiol 19: 360–367.

5. FlissPM, BruneW (2012) Prevention of cellular suicide by cytomegaloviruses. Viruses 4: 1928–1949.

6. RossiniG, CerboniC, SantoniA, LandiniMP, LandolfoS, et al. (2012) Interplay between human cytomegalovirus and intrinsic/innate host responses: a complex bidirectional relationship. Mediators Inflamm 2012: 607276.

7. SanchezV, SpectorDH (2008) Subversion of cell cycle regulatory pathways. Curr Top Microbiol Immunol 325: 243–262.

8. HocheggerH, TakedaS, HuntT (2008) Cyclin-dependent kinases and cell-cycle transitions: does one fit all? Nat Rev Mol Cell Biol 9: 910–916.

9. OhtsuboM, TheodorasAM, SchumacherJ, RobertsJM, PaganoM (1995) Human cyclin E, a nuclear protein essential for the G1-to-S phase transition. Mol Cell Biol 15: 2612–2624.

10. MailandN, DiffleyJF (2005) CDKs promote DNA replication origin licensing in human cells by protecting Cdc6 from APC/C-dependent proteolysis. Cell 122: 915–926.

11. KoeppDM, SchaeferLK, YeX, KeyomarsiK, ChuC, et al. (2001) Phosphorylation-dependent ubiquitination of cyclin E by the SCFFbw7 ubiquitin ligase. Science 294: 173–177.

12. StrohmaierH, SpruckCH, KaiserP, WonKA, SangfeltO, et al. (2001) Human F-box protein hCdc4 targets cyclin E for proteolysis and is mutated in a breast cancer cell line. Nature 413: 316–322.

13. Van LeukenR, ClijstersL, WolthuisR (2008) To cell cycle, swing the APC/C. Biochim Biophys Acta 1786: 49–59.

14. FurunoN, den ElzenN, PinesJ (1999) Human cyclin A is required for mitosis until mid prophase. J Cell Biol 147: 295–306.

15. GirardF, StrausfeldU, FernandezA, LambNJ (1991) Cyclin A is required for the onset of DNA replication in mammalian fibroblasts. Cell 67: 1169–79.

16. PaganoM, PepperkokR, VerdeF, AnsorgeW, DraettaG (1992) Cyclin A is required at two points in the human cell cycle. EMBO J 11: 961–971.

17. ResnitzkyD, HengstL, ReedSI (1995) Cyclin A-associated kinase activity is rate limiting for entrance into S phase and is negatively regulated in G1 by p27Kip1. Mol Cell Biol 15: 4347–4352.

18. ZindyF, LamasE, ChenivesseX, SobczakJ, WangJ, et al. (1992) Cyclin A is required in S phase in normal epithelial cells. Biochem Biophys Res Commun 182: 1144–1154.

19. De BoerL, OakesV, BeamishH, GilesN, StevensF, et al. (2008) Cyclin A/cdk2 coordinates centrosomal and nuclear mitotic events. Oncogene 27: 4261–4268.

20. FungTK, MaHT, PoonRY (2007) Specialized roles of the two mitotic cyclins in somatic cells: cyclin A as an activator of M phase-promoting factor. Mol Biol Cell 18: 1861–1873.

21. GongD, FerrellJEJr (2010) The roles of cyclin A2, B1, and B2 in early and late mitotic events. Mol Biol Cell 21: 3149–3161.

22. GongD, PomereningJR, MyersJW, GustavssonC, JonesJT, et al. (2007) Cyclin A2 regulates nuclear-envelope breakdown and the nuclear accumulation of cyclin B1. Curr Biol 17: 85–91.

23. LindqvistA, Rodriguez-BravoV, MedemaRH (2009) The decision to enter mitosis: feedback and redundancy in the mitotic entry network. J Cell Biol 185: 193–202.

24. MitraJ, EndersGH (2004) Cyclin A/Cdk2 complexes regulate activation of Cdk1 and Cdc25 phosphatases in human cells. Oncogene 23: 3361–3367.

25. BassermannF, von KlitzingC, MunchS, BaiRY, KawaguchiH, et al. (2005) NIPA defines an SCF-type mammalian E3 ligase that regulates mitotic entry. Cell 122: 45–57.

26. Lara-GonzalezP, WesthorpeFG, TaylorSS (2012) The spindle assembly checkpoint. Curr Biol 22: R966–980.

27. DirilMK, RatnacaramCK, PadmakumarVC, DuT, WasserM, et al. (2012) Cyclin-dependent kinase 1 (Cdk1) is essential for cell division and suppression of DNA re-replication but not for liver regeneration. Proc Natl Acad Sci U S A 109: 3826–3831.

28. KalaszczynskaI, GengY, IinoT, MizunoS, ChoiY, et al. (2009) Cyclin A is redundant in fibroblasts but essential in hematopoietic and embryonic stem cells. Cell 138: 352–365.

29. GopinathanL, TanSL, PadmakumarVC, CoppolaV, TessarolloL, et al. (2014) Loss of Cdk2 and Cyclin A2 Impairs Cell Proliferation and Tumorigenesis. Cancer Res 74: 3870–9 doi:10.1158/0008-5472

30. PagliucaFW, CollinsMO, LichawskaA, ZegermanP, ChoudharyJS, et al. (2011) Quantitative proteomics reveals the basis for the biochemical specificity of the cell-cycle machinery. Mol Cell 43: 406–417.

31. BresnahanWA, BoldoghI, ThompsonEA, AlbrechtT (1996) Human cytomegalovirus inhibits cellular DNA synthesis and arrests productively infected cells in late G1. Virology 224: 150–160.

32. HumeAJ, FinkelJS, KamilJP, CoenDM, CulbertsonMR, et al. (2008) Phosphorylation of retinoblastoma protein by viral protein with cyclin-dependent kinase function. Science 320: 797–799.

33. KalejtaRF, ShenkT (2003) Proteasome-dependent, ubiquitin-independent degradation of the Rb family of tumor suppressors by the human cytomegalovirus pp71 protein. Proc Natl Acad Sci U S A 100: 3263–3268.

34. SongYJ, StinskiMF (2002) Effect of the human cytomegalovirus IE86 protein on expression of E2F-responsive genes: a DNA microarray analysis. Proc Natl Acad Sci U S A 99: 2836–2841.

35. BresnahanWA, AlbrechtT, ThompsonEA (1998) The cyclin E promoter is activated by human cytomegalovirus 86-kDa immediate early protein. J Biol Chem 273: 22075–22082.

36. BresnahanWA, ThompsonEA, AlbrechtT (1997) Human cytomegalovirus infection results in altered Cdk2 subcellular localization. J Gen Virol 78 (Pt 8): 1993–1997.

37. McElroyAK, DwarakanathRS, SpectorDH (2000) Dysregulation of cyclin E gene expression in human cytomegalovirus-infected cells requires viral early gene expression and is associated with changes in the Rb-related protein p130. J Virol 74: 4192–4206.

38. WiebuschL, HagemeierC (2001) The human cytomegalovirus immediate early 2 protein dissociates cellular DNA synthesis from cyclin-dependent kinase activation. EMBO J 20: 1086–1098.

39. BiswasN, SanchezV, SpectorDH (2003) Human cytomegalovirus infection leads to accumulation of geminin and inhibition of the licensing of cellular DNA replication. J Virol 77: 2369–2376.

40. QianZ, Leung-PinedaV, XuanB, Piwnica-WormsH, YuD (2010) Human cytomegalovirus protein pUL117 targets the mini-chromosome maintenance complex and suppresses cellular DNA synthesis. PLoS Pathog 6: e1000814.

41. WiebuschL, UeckerR, HagemeierC (2003) Human cytomegalovirus prevents replication licensing by inhibiting MCM loading onto chromatin. EMBO Rep 4: 42–46.

42. FehrAR, GualbertoNC, SavarynJP, TerhuneSS, YuD (2012) Proteasome-dependent disruption of the E3 ubiquitin ligase anaphase-promoting complex by HCMV protein pUL21a. PLoS Pathog 8: e1002789.

43. TranK, KamilJP, CoenDM, SpectorDH (2010) Inactivation and disassembly of the anaphase-promoting complex during human cytomegalovirus infection is associated with degradation of the APC5 and APC4 subunits and does not require UL97-mediated phosphorylation of Cdh1. J Virol 84: 10832–10843.

44. WiebuschL, BachM, UeckerR, HagemeierC (2005) Human cytomegalovirus inactivates the G0/G1-APC/C ubiquitin ligase by Cdh1 dissociation. Cell Cycle 4: 1435–1439.

45. JaultFM, JaultJM, RuchtiF, FortunatoEA, ClarkC, et al. (1995) Cytomegalovirus infection induces high levels of cyclins, phosphorylated Rb, and p53, leading to cell cycle arrest. J Virol 69: 6697–6704.

46. ShlapoberskyM, SandersR, ClarkC, SpectorDH (2006) Repression of HMGA2 gene expression by human cytomegalovirus involves the IE2 86-kilodalton protein and is necessary for efficient viral replication and inhibition of cyclin A transcription. J Virol 80: 9951–9961.

47. SanchezV, McElroyAK, SpectorDH (2003) Mechanisms governing maintenance of Cdk1/cyclin B1 kinase activity in cells infected with human cytomegalovirus. J Virol 77: 13214–13224.

48. BainM, SinclairJ (2007) The S phase of the cell cycle and its perturbation by human cytomegalovirus. Rev Med Virol 17: 423–434.

49. O'DowdJM, ZavalaAG, BrownCJ, MoriT, FortunatoEA (2012) HCMV-infected cells maintain efficient nucleotide excision repair of the viral genome while abrogating repair of the host genome. PLoS Pathog 8: e1003038.

50. SalvantBS, FortunatoEA, SpectorDH (1998) Cell cycle dysregulation by human cytomegalovirus: influence of the cell cycle phase at the time of infection and effects on cyclin transcription. J Virol 72: 3729–3741.

51. OduroJD, UeckerR, HagemeierC, WiebuschL (2012) Inhibition of human cytomegalovirus immediate-early gene expression by cyclin A2-dependent kinase activity. J Virol 86: 9369–9383.

52. ZydekM, HagemeierC, WiebuschL (2010) Cyclin-dependent kinase activity controls the onset of the HCMV lytic cycle. PLoS Pathog 6: e1001096.

53. BogdanowB, WeisbachH, von EinemJ, StraschewskiS, VoigtS, et al. (2013) Human cytomegalovirus tegument protein pp150 acts as a cyclin A2-CDK-dependent sensor of the host cell cycle and differentiation state. Proc Natl Acad Sci U S A 110: 17510–17515.

54. BrownNR, NobleME, EndicottJA, JohnsonLN (1999) The structural basis for specificity of substrate and recruitment peptides for cyclin-dependent kinases. Nat Cell Biol 1: 438–443.

55. FehrAR, YuD (2010) Human cytomegalovirus gene UL21a encodes a short-lived cytoplasmic protein and facilitates virus replication in fibroblasts. J Virol 84: 291–302.

56. Stern-GinossarN, WeisburdB, MichalskiA, LeVT, HeinMY, et al. (2012) Decoding human cytomegalovirus. Science 338: 1088–1093.

57. LoweED, TewsI, ChengKY, BrownNR, GulS, et al. (2002) Specificity determinants of recruitment peptides bound to phospho-CDK2/cyclin A. Biochemistry 41: 15625–15634.

58. SchulmanBA, LindstromDL, HarlowE (1998) Substrate recruitment to cyclin-dependent kinase 2 by a multipurpose docking site on cyclin A. Proc Natl Acad Sci U S A 95: 10453–10458.

59. BrownNR, LoweED, PetriE, SkamnakiV, AntrobusR, et al. (2007) Cyclin B and cyclin A confer different substrate recognition properties on CDK2. Cell Cycle 6: 1350–1359.

60. PenfoldME, MocarskiES (1997) Formation of cytomegalovirus DNA replication compartments defined by localization of viral proteins and DNA synthesis. Virology 239: 46–61.

61. StrangBL, BoulantS, ChangL, KnipeDM, KirchhausenT, et al. (2012) Human cytomegalovirus UL44 concentrates at the periphery of replication compartments, the site of viral DNA synthesis. J Virol 86: 2089–2095.

62. StenbergK, SkogS, TribukaitB (1985) Concentration-dependent effects of foscarnet on the cell cycle. Antimicrob Agents Chemother 28: 802–806.

63. GavetO, PinesJ (2010) Progressive activation of CyclinB1-Cdk1 coordinates entry to mitosis. Dev Cell 18: 533–543.

64. StevensJB, AbdallahBY, ReganSM, LiuG, BremerSW, et al. (2010) Comparison of mitotic cell death by chromosome fragmentation to premature chromosome condensation. Mol Cytogenet 3: 20.

65. El AchkarE, Gerbault-SeureauM, MulerisM, DutrillauxB, DebatisseM (2005) Premature condensation induces breaks at the interface of early and late replicating chromosome bands bearing common fragile sites. Proc Natl Acad Sci U S A 102: 18069–18074.

66. GorbskyGJ (2013) Cohesion fatigue. Curr Biol 23: R986–8.

67. AhnJH, JangWJ, HaywardGS (1999) The human cytomegalovirus IE2 and UL112–113 proteins accumulate in viral DNA replication compartments that initiate from the periphery of promyelocytic leukemia protein-associated nuclear bodies (PODs or ND10). J Virol 73: 10458–10471.

68. LafeminaRL, PizzornoMC, MoscaJD, HaywardGS (1989) Expression of the acidic nuclear immediate-early protein (IE1) of human cytomegalovirus in stable cell lines and its preferential association with metaphase chromosomes. Virology 172: 584–600.

69. MückeK, PaulusC, BernhardtK, GerrerK, SchönK, et al. (2014) Human cytomegalovirus major immediate early 1 protein targets host chromosomes by docking to the acidic pocket on the nucleosome surface. J Virol 88: 1228–1248.

70. FehrAR, YuD (2013) Control the host cell cycle: viral regulation of the anaphase-promoting complex. J Virol 87: 8818–8825.

71. WooRA, PoonRY (2003) Cyclin-dependent kinases and S phase control in mammalian cells. Cell Cycle 2: 316–324.

72. KatsunoY, SuzukiA, SugimuraK, OkumuraK, ZineldeenDH, et al. (2009) Cyclin A-Cdk1 regulates the origin firing program in mammalian cells. Proc Natl Acad Sci U S A 106: 3184–3189.

73. PooleE, BainM, TeagueL, TakeiY, LaskeyR, et al. (2012) The cellular protein MCM3AP is required for inhibition of cellular DNA synthesis by the IE86 protein of human cytomegalovirus. PLoS One 7: e45686.

74. IshimiY, Komamura-KohnoY, YouZ, OmoriA, KitagawaM (2000) Inhibition of Mcm4,6,7 helicase activity by phosphorylation with cyclin A/Cdk2. J Biol Chem 275: 16235–16241.

75. MoritaniM, IshimiY (2013) Inhibition of DNA binding of MCM2-7 complex by phosphorylation with cyclin-dependent kinases. J Biochem 154: 363–372.

76. WheelerLW, LentsNH, BaldassareJJ (2008) Cyclin A-CDK activity during G1 phase impairs MCM chromatin loading and inhibits DNA synthesis in mammalian cells. Cell Cycle 7: 2179–2188.

77. ChibazakuraT, KamachiK, OharaM, TaneS, YoshikawaH, et al. (2011) Cyclin A promotes S-phase entry via interaction with the replication licensing factor Mcm7. Mol Cell Biol 31: 248–255.

78. FehrAR, YuD (2011) Human cytomegalovirus early protein pUL21a promotes efficient viral DNA synthesis and the late accumulation of immediate-early transcripts. J Virol 85: 663–674.

79. CaffarelliN, FehrAR, YuD (2013) Cyclin A degradation by primate cytomegalovirus protein pUL21a counters its innate restriction of virus replication. PLoS Pathog 9: e1003825.

80. WiebuschL, NeuwirthA, GrabenhenrichL, VoigtS, HagemeierC (2008) Cell cycle-independent expression of immediate-early gene 3 results in G1 and G2 arrest in murine cytomegalovirus-infected cells. J Virol 82: 10188–10198.

81. BoldoghI, AbuBakarS, AlbrechtT (1990) Activation of proto-oncogenes: an immediate early event in human cytomegalovirus infection. Science 247: 561–564.

82. HertelL, MocarskiES (2004) Global analysis of host cell gene expression late during cytomegalovirus infection reveals extensive dysregulation of cell cycle gene expression and induction of pseudomitosis independent of US28 function. J Virol 78: 11988–12011.

83. BartkovaJ, HorejsíZ, KoedK, KrämerA, TortF, et al. (2005) DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434: 864–870.

84. CanmanCE (2001) Replication checkpoint: preventing mitotic catastrophe. Curr Biol 11: R121–124.

85. GasparM, ShenkT (2006) Human cytomegalovirus inhibits a DNA damage response by mislocalizing checkpoint proteins. Proc Natl Acad Sci U S A 103: 2821–2826.

86. SavarynJP, ReitsmaJM, BigleyTM, HalliganBD, QianZ, et al. (2013) Human cytomegalovirus pUL29/28 and pUL38 repression of p53-regulated p21CIP1 and caspase 1 promoters during infection. J Virol 87: 2463–2474.

87. XiaofeiE, PickeringMT, DebatisM, CastilloJ, LagadinosA, et al. (2011) An E2F1-mediated DNA damage response contributes to the replication of human cytomegalovirus. PLoS Pathog 7: e1001342.

88. XiaofeiE, SavidisG, ChinCR, WangS, LuS, et al. (2014) A Novel DDB2-ATM Feedback Loop Regulates Human Cytomegalovirus Replication. J Virol 88: 2279–2290.

89. BeeharryN, RattnerJB, CavistonJP, YenT (2013) Centromere fragmentation is a common mitotic defect of S and G2 checkpoint override. Cell Cycle 12: 1588–1597.

90. Del NagroCJ, ChoiJ, XiaoY, RangellL, MohanS, et al. (2014) Chk1 inhibition in p53 deficient cell lines drives rapid chromosome fragmentation followed by caspase-independent cell death. Cell Cycle 13: 303–314.

91. GanemNJ, PellmanD (2012) Linking abnormal mitosis to the acquisition of DNA damage. J Cell Biol 199: 871–881.

92. DaumJR, PotapovaTA, SivakumarS, DanielJJ, FlynnJN, et al. (2011) Cohesion fatigue induces chromatid separation in cells delayed at metaphase. Curr Biol 21: 1018–1024.

93. StevensD, GassmannR, OegemaK, DesaiA (2011) Uncoordinated loss of chromatid cohesion is a common outcome of extended metaphase arrest. PLoS One 6: e22969.

94. MichaelisM, DoerrHW, CinatlJ (2009) The story of human cytomegalovirus and cancer: increasing evidence and open questions. Neoplasia 11: 1–9.

95. DziurzynskiK, ChangSM, HeimbergerAB, KalejtaRF, McGregor DallasSR, et al. (2012) HCMV and Gliomas Symposium. Consensus on the role of human cytomegalovirus in glioblastoma. Neuro Oncol 14: 246–255.

96. SinzgerC, HahnG, DigelM, KatonaR, SampaioKL, et al. (2008) Cloning and sequencing of a highly productive, endotheliotropic virus strain derived from human cytomegalovirus TB40/E. J Gen Virol 89: 359–368.

97. TischerBK, SmithGA, OsterriederN (2010) En passant mutagenesis: a two step markerless red recombination system. Methods Mol Biol 634: 421–430.

98. SaffertRT, KalejtaRF (2007) Human cytomegalovirus gene expression is silenced by Daxx-mediated intrinsic immune defense in model latent infections established in vitro. J Virol 81: 9109–9120.