Ubiquitination and sumoylation of the HTLV-2 Tax-2B protein regulate its NF-κB activity: a comparative study with the HTLV-1 Tax-1 protein

Retrovirology

Maria Grazia Romanelli³  Umberto Bertazzoni³  Françoise Bex¹  Carla Sampaio¹  Erica Diani³  Oriano Marin²  Anne Sophie Rinaldi¹  Gianfranco Di Gennaro³  Julie Lodewick¹  Marco Turci³ 

[1] Institute for Microbiological Research J-M Wiame, Laboratory of Microbiology, Université Libre de Bruxelles, 1, Avenue E. Gryson, Brussels, Belgium;Department of Oncology and Surgical Sciences, University of Padova, Padova, Italy;Department of Life and Reproduction Sciences, Section of Biology and Genetics, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy

关键词: NF-κB pathway;    Sumoylation;    Ubiquitination;    Post-translational modification;    Leukemia;    Oncoprotein;    Tax;    Retrovirus;    HTLV-2;    HTLV-1;   

Others  :  1209222
DOI  :  10.1186/1742-4690-9-102

 received in 2012-04-18, accepted in 2012-11-02,  发布年份 2012

【 摘 要 】

Background

Retroviruses HTLV-1 and HTLV-2 have homologous genomic structures but differ significantly in pathogenicity. HTLV-1 is associated with Adult T cell Leukemia (ATL), whereas infection by HTLV-2 has no association with neoplasia. Transformation of T lymphocytes by HTLV-1 is linked to the capacity of its oncoprotein Tax-1 to alter cell survival and cell cycle control mechanisms. Among these functions, Tax-1-mediated activation of cellular gene expression via the NF-κB pathway depends on Tax-1 post-translational modifications by ubiquitination and sumoylation. The Tax-2 protein of HTLV-2B (Tax-2B) is also modified by ubiquitination and sumoylation and activates the NF-κB pathway to a level similar to that of Tax-1. The present study aims to understand whether ubiquitination and sumoylation modifications are involved in Tax-2B-mediated activation of the NF-κB pathway.

Results

The comparison of Tax-1 and Tax-2B lysine to arginine substitution mutants revealed conserved patterns and levels of ubiquitination with notable difference in the lysine usage for sumoylation. Neither Tax-1 nor Tax-2B ubiquitination and sumoylation deficient mutants could activate the NF-κB pathway and fusion of ubiquitin or SUMO-1 to the C-terminus of the ubiquitination and sumoylation deficient Tax-2B mutant strikingly restored transcriptional activity. In addition, ubiquitinated forms of Tax-2B colocalized with RelA and IKKγ in prominent cytoplasmic structures associated with the Golgi apparatus, whereas colocalization of Tax-2B with the RelA subunit of NF-κB and the transcriptional coactivator p300 in punctate nuclear structures was dependent on Tax-2B sumoylation, as previously observed for Tax-1.

Conclusions

Both Tax-1 and Tax-2 activate the NF-κB pathway via similar mechanisms involving ubiquitination and sumoylation. Therefore, the different transforming potential of HTLV-1 and HTLV-2 is unlikely to be related to different modes of activation of the canonical NF-κB pathway.

【 授权许可】

   
2012 Turci et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Matsuoka M, Jeang KT: Human T-cell leukaemia virus type 1 (HTLV-1) infectivity and cellular transformation. Nat Rev Cancer 2007, 7:270-280.
• [2]Araujo A, Hall WW: Human T-lymphotropic virus type II and neurological disease. Ann Neurol 2004, 56:10-19.
• [3]Lewis MJ, Sheehy N, Salemi M, Vandamme AM, Hall WW: Comparison of CREB- and NF-kappaB-mediated transactivation by human T lymphotropic virus type II (HTLV-II) and type I (HTLV-I) tax proteins. Virology 2002, 295:182-189.
• [4]Grassmann R, Aboud M, Jeang KT: Molecular mechanisms of cellular transformation by HTLV-1 Tax. Oncogene 2005, 24:5976-5985.
• [5]Marriott SJ, Semmes OJ: Impact of HTLV-I Tax on cell cycle progression and the cellular DNA damage repair response. Oncogene 2005, 24:5986-5995.
• [6]Giam CZ, Jeang KT: HTLV-1 Tax and adult T-cell leukemia. Front Biosci 2007, 12:1496-1507.
• [7]Boxus M, Willems L: Mechanisms of HTLV-1 persistence and transformation. Br J Cancer 2009, 101:1497-1501.
• [8]Boxus M, Willems L: How the DNA damage response determines the fate of HTLV-1 Tax-expressing cells. Retrovirology 2012, 9:2. BioMed Central Full Text
• [9]Zane L, Sibon D, Jeannin L, Zandecki M, Delfau-Larue MH, Gessain A, et al.: Tax gene expression and cell cycling but not cell death are selected during HTLV-1 infection in vivo. Retrovirology 2010, 7:17. BioMed Central Full Text
• [10]Mahieux R, Pise-Masison CA, Nicot C, Green P, Hall WW, Brady JN: Inactivation of p53 by HTLV type 1 and HTLV type 2 Tax trans-activators. AIDS Res Hum Retroviruses 2000, 16:1677-1681.
• [11]Peloponese JM Jr, Kinjo T, Jeang KT: Human T-cell leukemia virus type 1 Tax and cellular transformation. Int J Hematol 2007, 86:101-106.
• [12]Bertazzoni U, Turci M, Avesani F, di Gennaro G, Bidoia C, Romanelli MG: Intracellular localization and cellular factors interaction of HTLV-1 and HTLV-2 Tax proteins: similarities and functional differences. Viruses 2011, 3:541-560.
• [13]Higuchi M, Tsubata C, Kondo R, Yoshida S, Takahashi M, Oie M, et al.: Cooperation of NF-kappaB2/p100 activation and the PDZ domain binding motif signal in human T-cell leukemia virus type 1 (HTLV-1) Tax1 but not HTLV-2 Tax2 is crucial for interleukin-2-independent growth transformation of a T-cell line. J Virol 2007, 81:11900-11907.
• [14]Shoji T, Higuchi M, Kondo R, Takahashi M, Oie M, Tanaka Y, et al.: Identification of a novel motif responsible for the distinctive transforming activity of human T-cell leukemia virus (HTLV) type 1 Tax1 protein from HTLV-2 Tax2. Retrovirology 2009, 6:83. BioMed Central Full Text
• [15]Semmes OJ, Majone F, Cantemir C, Turchetto L, Hjelle B, Jeang KT: HTLV-I and HTLV-II Tax: differences in induction of micronuclei in cells and transcriptional activation of viral LTRs. Virology 1996, 217:373-379.
• [16]Hirata A, Higuchi M, Niinuma A, Ohashi M, Fukushi M, Oie M, et al.: PDZ domain-binding motif of human T-cell leukemia virus type 1 Tax oncoprotein augments the transforming activity in a rat fibroblast cell line. Virology 2004, 318:327-336.
• [17]Rousset R, Fabre S, Desbois C, Bantignies F, Jalinot P: The C-terminus of the HTLV-1 Tax oncoprotein mediates interaction with the PDZ domain of cellular proteins. Oncogene 1998, 16:643-654.
• [18]Xie L, Yamamoto B, Haoudi A, Semmes OJ, Green PL: PDZ binding motif of HTLV-1 Tax promotes virus-mediated T-cell proliferation in vitro and persistence in vivo. Blood 2006, 107:1980-1988.
• [19]Van PL, Yim KW, Jin DY, Dapolito G, Kurimasa A, Jeang KT: Genetic evidence of a role for ATM in functional interaction between human T-cell leukemia virus type 1 Tax and p53. J Virol 2001, 75:396-407.
• [20]Lodewick J, Lamsoul I, Bex F: Move or die: the fate of the Tax oncoprotein of HTLV-1. Viruses 2011, 3:829-857.
• [21]Kfoury Y, Nasr R, Journo C, Mahieux R, Pique C, Bazarbachi A: The multifaceted oncoprotein Tax: subcellular localization, posttranslational modifications, and NF-kappaB activation. Adv Cancer Res 2012, 113:85-120.
• [22]Lamsoul I, Lodewick J, Lebrun S, Brasseur R, Burny A, Gaynor RB, et al.: Exclusive ubiquitination and sumoylation on overlapping lysine residues mediate NF-kappaB activation by the human T-cell leukemia virus tax oncoprotein. Mol Cell Biol 2005, 25:10391-10406.
• [23]Harhaj NS, Sun SC, Harhaj EW: Activation of NF-kappa B by the human T cell leukemia virus type I Tax oncoprotein is associated with ubiquitin-dependent relocalization of I kappa B kinase. J Biol Chem 2007, 282:4185-4192.
• [24]Shembade N, Harhaj NS, Yamamoto M, Akira S, Harhaj EW: The human T-cell leukemia virus type 1 Tax oncoprotein requires the ubiquitin-conjugating enzyme Ubc13 for NF-kappaB activation. J Virol 2007, 81:13735-13742.
• [25]Nasr R, Chiari E, El Sabban M, Mahieux R, Kfoury Y, Abdulhay M, et al.: Tax ubiquitylation and sumoylation control critical cytoplasmic and nuclear steps of NF-kappa B activation. Blood 2006, 107:4021-4029.
• [26]Bex F, McDowall A, Burny A, Gaynor R: The human T-cell leukemia virus type 1 transactivator protein Tax colocalizes in unique nuclear structures with NF-kappaB proteins. J Virol 1997, 71:3484-3497.
• [27]Bex F, Yin MJ, Burny A, Gaynor RB: Differential transcriptional activation by human T-cell leukemia virus type 1 Tax mutants is mediated by distinct interactions with CREB binding protein and p300. Mol Cell Biol 1998, 18:2392-2405.
• [28]Kfoury Y, Setterblad N, El Sabban M, Zamborlini A, Dassouki Z, El Hajj H, et al.: Tax ubiquitylation and SUMOylation control the dynamic shuttling of Tax and NEMO between Ubc9 nuclear bodies and the centrosome. Blood 2011, 117:190-199.
• [29]Turci M, Lodewick J, Righi P, Polania A, Romanelli MG, Bex F, et al.: HTLV-2B Tax oncoprotein is modified by ubiquitination and sumoylation and displays intracellular localization similar to its homologue HTLV-1 Tax. Virology 2009, 386:6-11.
• [30]Avesani F, Romanelli MG, Turci M, di Gennaro G, Sampaio C, Bidoia C, et al.: Association of HTLV Tax proteins with TAK1-binding protein 2 and RelA in calreticulin-containing cytoplasmic structures participates in Tax-mediated NF-kappaB activation. Virology 2010, 408:39-48.
• [31]Meertens L, Chevalier S, Weil R, Gessain A, Mahieux R: A 10-amino acid domain within human T-cell leukemia virus type 1 and type 2 tax protein sequences is responsible for their divergent subcellular distribution. J Biol Chem 2004, 279:43307-43320.
• [32]Wertz IE, Dixit VM: Signaling to NF-kappaB: regulation by ubiquitination. Cold Spring Harb Perspect Biol 2010, 2:a003350.
• [33]Gohda J, Irisawa M, Tanaka Y, Sato S, Ohtani K, Fujisawa J, et al.: HTLV-1 Tax-induced NFkappaB activation is independent of Lys-63-linked-type polyubiquitination. Biochem Biophys Res Commun 2007, 357:225-230.
• [34]Van Damme E, Laukens K, Dang TH, Van Ostade X: A manually curated network of the PML nuclear body interactome reveals an important role for PML-NBs in SUMOylation dynamics. Int J Biol Sci 2010, 6:51-67.
• [35]Bonnet A, Randrianarison-Huetz V, Nzounza P, Nedelec M, Chazal M, Waast L, et al.: Low nuclear body formation and tax SUMOylation do not prevent NF-kappaB promoter activation. Retrovirology 2012, 9:77. BioMed Central Full Text
• [36]Hay RT: SUMO: a history of modification. Mol Cell 2005, 18:1-12.
• [37]Iborra FJ, Pombo A, McManus J, Jackson DA, Cook PR: The topology of transcription by immobilized polymerases. Exp Cell Res 1996, 229:167-173.
• [38]Xiao G: NF-kappaB activation: Tax sumoylation is out, but what about Tax ubiquitination? Retrovirology 2012, 9:78. BioMed Central Full Text
• [39]Takebe Y, Seiki M, Fujisawa J, Hoy P, Yokota K, Arai K, et al.: SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol Cell Biol 1988, 8:466-472.
• [40]Chiari E, Lamsoul I, Lodewick J, Chopin C, Bex F, Pique C: Stable ubiquitination of human T-cell leukemia virus type 1 tax is required for proteasome binding. J Virol 2004, 78:11823-11832.
• [41]Tatham MH, Jaffray E, Vaughan OA, Desterro JM, Botting CH, Naismith JH, et al.: Polymeric chains of SUMO-2 and SUMO-3 are conjugated to protein substrates by SAE1/SAE2 and Ubc9. J Biol Chem 2001, 276:35368-35374.
• [42]Hanstein B, Eckner R, DiRenzo J, Halachmi S, Liu H, Searcy B, et al.: p300 Is a component of an Estrogen receptor coactivator complex. Proc Natl Acad Sci U S A 1996, 93:11540-11545.