[en] Bovine leukemia virus (BLV) is a deltaretrovirus infecting bovine B cells and causing enzootic bovine leucosis (EBL). The long terminal repeat (LTR) plays an indispensable role in viral gene expression. The BLV Tax protein acts as the main transactivator of LTR-driven transcription of BLV viral genes. The aim of this study was to analyze mutations in the BLV LTR region and tax gene to determine their association with transcriptional activity. LTRs were obtained from one hundred and six BLV isolates and analyzed for their genetic variability. Fifteen variants were selected and characterized based on mutations in LTR regulatory elements, and further used for in vitro transcription assays. Reporter vectors containing the luciferase gene under the control of each
variant BLV promoter sequence, in addition to variant Tax expression vectors, were constructed. Both types of plasmids were used for cotransfection of HeLa cells and the level of luciferase activity was measured as a proxy of transcriptional activity. Marked differences in LTR promoter activity
and Tax transactivation activity were observed amongst BLV variants. These results demonstrate that mutations in both the BLV LTR and tax gene can affect the promoter activity, which may have important consequences on proviral load, viral fitness, and transmissibility in BLV-infected cattle.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Pluta, Aneta; National Veterinary Research Institute, Puławy
Willems, Luc ; Université de Liège - ULiège > Cancer-Cellular and Molecular Epigenetics
Douville, Renée N.
Kuzmak, Jacek; National Veterinary Research Institute, Puławy
Language :
English
Title :
Effects of Naturally Occurring Mutations in Bovine Leukemia Virus 50 -LTR and Tax Gene on Viral Transcriptional Activity
Publication date :
13 October 2020
Journal title :
Pathogens
eISSN :
2076-0817
Publisher :
Molecular Diversity Preservation International (MDPI), Basel, Switzerland
Volume :
9
Issue :
10
Pages :
836
Peer reviewed :
Peer Reviewed verified by ORBi
Funders :
NCN - Narodowe Centrum Nauki [PL] Fund for Scientific Research at the National Veterinary Research Institute, Puławy, Poland
Walker, P.J.; Siddell, S.G.; Lefkowitz, E.J.; Mushegian, A.R.; Dempsey, D.M.; Dutilh, B.E.; Harrach, B.; Harrison, R.L.; Hendrickson, R.C.; Junglen, S.; et al. Changes to virus taxonomy and the International Code of Virus Classification and Nomenclature ratified by the International Committee on Taxonomy of Viruses (2019). Arch. Virol. 2019, 164, 2417–2429.
Rodríguez, S.M.; Florins, A.; Gillet, N.A.; De Brogniez, A.; Sánchez‐Alcaraz, M.T.; Boxus, M.; Boulanger, F.; Gutierrez, G.; Trono, K.; Alvarez, I.; et al. Preventive and Therapeutic Strategies for Bovine Leukemia Virus: Lessons for HTLV. Viruses 2011, 3, 1210–1248.
Murakami, K.; Kobayashi, S.; Konishi, M.; Kameyama, K.; Yamamoto, T.; Tsutsui, T. The recent prevalence of bovine leukemia virus (BLV) infection among Japanese cattle. Vet. Microbiol. 2011, 148, 84–88.
LaDronka, R.M.; Ainsworth, S.; Wilkins, M.J.; Norby, B.; Byrem, T.M.; Bartlett, P.C. Prevalence of Bovine Leukemia Virus Antibodies in US Dairy Cattle. Vet. Med. Int. 2018, 2018, 5831278.
Rola‐Luszczak, M.; Pluta, A.; Olech, M.; Donnik, I.; Petropavlovskiy, M.; Gerilovych, A.; Vinogradova, I.; Choudhury, B.; Kuzmak, J. The molecular characterization of bovine leukaemia virus isolates from Eastern Europe and Siberia and its impact on phylogeny. PLoS ONE 2013, 8, e58705.
Sagata, N.; Yasunaga, T.; Ogawa, Y.; Tsuzuku‐Kawamura, J.; Ikawa, Y. Bovine leukemia virus: Unique structural features of its long terminal repeats and its evolutionary relationship to human T‐cell leukemia virus. Proc. Natl. Acad. Sci. USA 1984, 81, 4741–4745.
Kettmann, R.; Cleuter, Y.; Gregoire, D.; Burny, A. Role of the 3′ long open reading frame region of bovine leukemia virus in the maintenance of cell transformation. J. Virol. 1985, 54, 899–901.
Gillet, N.; Florins, A.; Boxus, M.; Burteau, C.; Nigro, A.; Vandermeers, F.; Balon, H.; Bouzar, A.B.; Defoiche, J.; Burny, A.; et al. Mechanisms of leukemogenesis induced by bovine leukemia virus: Prospects for novel anti‐retroviral therapies in human. Retrovirology 2007, 4, 18.
Kincaid, R.P.; Burke, J.M.; Sullivan, C.S. RNA virus microRNA that mimics a B‐cell oncomiR. Proc. Natl. Acad. Sci. USA 2012, 109, 3077–3082.
Rosewick, N.; Momont, M.; Durkin, K.; Takeda, H.; Caiment, F.; Cleuter, Y.; Vernin, C.; Mortreux, F.; Wattel, E.; Burny, A.; et al. Deep sequencing reveals abundant noncanonical retroviral microRNAs in B‐ cell leukemia/lymphoma. Proc. Natl. Acad. Sci. USA 2013, 110, 2306–2311.
Derse, D.; Casey, J.W. Two elements in the bovine leukemia virus long terminal repeat that regulate gene expression. Science 1986, 231, 1437–1440.
Couez, D.; Deschamps, J.; Kettmann, R.; Stephens, R.M.; Gilden, R.V.; Burny, A. Nucleotide sequence analysis of the long terminal repeat of integrated bovine leukemia provirus DNA and of adjacent viral and host sequences. J. Virol. 1984, 49, 615–620.
Katoh, I.; Yoshinaka, Y.; Ikawa, Y. Bovine leukemia virus trans‐activator p38tax activates heterologous promoters with a common sequence known as a cAMP‐responsive element or the binding site of a cellular transcription factor ATF. EMBO J. 1989, 8, 497–503.
Merezak, C.; Pierreux, C.; Adam, E.; Lemaigre, F.; Rousseau, G.G.; Calomme, C.; Van Lint, C.; Christophe, D.; Kerkhofs, P.; Burny, A.; et al. Suboptimal enhancer sequences are required for efficient bovine leukemia virus propagation in vivo: Implications for viral latency. J. Virol. 2001, 75, 6977–6988.
Boros, I.M.; Tie, F.; Giam, C.Z.Interaction of bovine leukemia virus transactivator Tax with bZip proteins. Virology 1995, 214, 207–214.
Willems, L.; Kettmann, R.; Chen, G.; Portetelle, D.; Burny, A.; Derse, D. A cyclic AMP‐responsive DNA-binding protein (CREB2) is a cellular transactivator of the bovine leukemia virus long terminal repeat. J. Virol. 1992, 66, 766–772.
Brooks, P.A.; Nyborg, J.K.; Cockerell, G.L. Identification of an NF‐kappa B binding site in the bovine leukemia virus promoter. J. Virol. 1995, 69, 6005–6009.
Niermann, G.L.; Buehring, G.C. Hormone regulation of bovine leukemia virus via the long terminal repeat. Virology 1997, 239, 249–258.
Dekoninck, A.; Calomme, C.; Nizet, S.; de Launoit, Y.; Burny, A.; Ghysdael, J.; Van Lint, C. Identification and characterization of a PU.1/Spi‐B binding site in the bovine leukemia virus long terminal repeat. Oncogene 2003, 22, 2882–2896.
Calomme, C.; Nguyen, T.L.; de Launoit, Y.; Kiermer, V.; Droogmans, L.; Burny, A.; Van Lint, C. Upstream stimulatory factors binding to an E box motif in the R region of the bovine leukemia virus long terminal repeat stimulates viral gene expression. J. Biol. Chem. 2002, 277, 8775–8789.
Kiss‐Toth, E.; Unk, I. A downstream regulatory element activates the bovine leukemia virus promoter. Biochem. Biophys. Res. Commun. 1994, 202, 1553–1561.
Mansky, L.M.; Temin, H.M. Lower mutation rate of bovine leukemia virus relative to that of spleen necrosis virus. J. Virol. 1994, 68, 494–499.
Moratorio, G.; Fischer, S.; Bianchi, S.; Tome, L.; Rama, G.; Obal, G.; Carrion, F.; Pritsch, O.; Cristina, J. A detailed molecular analysis of complete bovine leukemia virus genomes isolated from B‐cell lymphosarcomas. Vet. Res. 2013, 44, 19.
Coulston, J.; Naif, H.; Brandon, R.; Kumar, S.; Khan, S.; Daniel, R.C.; Lavin, M.F. Molecular cloning and sequencing of an Australian isolate of proviral bovine leukaemia virus DNA: Comparison with other isolates. J. Gen. Virol. 1990, 71, 1737–1746.
Camargos, M.F.; Stancek, D.; Rocha, M.A.; Lessa, L.M.; Reis, J.K.; Leite, R.C. Partial sequencing of env gene of bovine leukaemia virus from Brazilian samples and phylogenetic analysis. J. Vet. Med. B Infect. Dis. Vet. Public Health 2002, 49, 325–331.
Camargos, M.F.; Pereda, A.; Stancek, D.; Rocha, M.A.; dos Reis, J.K.; Greiser‐Wilke, I.; Leite, R.C. Molecular characterization of the env gene from Brazilian field isolates of Bovine leukemia virus. Virus Genes 2007, 34, 343–350.
Mamoun, R.Z.; Morisson, M.; Rebeyrotte, N.; Busetta, B.; Couez, D.; Kettmann, R.; Hospital, M.; Guillemain, B. Sequence variability of bovine leukemia virus env gene and its relevance to the structure and antigenicity of the glycoproteins. J. Virol. 1990, 64, 4180–4188.
Fechner, H.; Blankenstein, P.; Looman, A.C.; Elwert, J.; Geue, L.; Albrecht, C.; Kurg, A.; Beier, D.; Marquardt, O.; Ebner, D. Provirus variants of the bovine leukemia virus and their relation to the serological status of naturally infected cattle. Virology 1997, 237, 261–269.
Balic, D.; Lojkic, I.; Periskic, M.; Bedekovic, T.; Jungic, A.; Lemo, N.; Roic, B.; Cac, Z.; Barbic, L.; Madic, J. Identification of a new genotype of bovine leukemia virus. Arch. Virol. 2012, 157, 1281–1290.
Polat, M.; Takeshima, S.N.; Hosomichi, K.; Kim, J.; Miyasaka, T.; Yamada, K.; Arainga, M.; Murakami, T.; Matsumoto, Y.; de la Barra Diaz, V.; et al. A new genotype of bovine leukemia virus in South America identified by NGS‐based whole genome sequencing and molecular evolutionary genetic analysis. Retrovirology 2016, 13, 4.
Polat, M.; Moe, H.H.; Shimogiri, T.; Moe, K.K.; Takeshima, S.N.; Aida, Y. The molecular epidemiological study of bovine leukemia virus infection in Myanmar cattle. Arch. Virol. 2017, 162, 425–437.
Murakami, H.; Todaka, H.; Uchiyama, J.; Sato, R.; Sogawa, K.; Sakaguchi, M.; Tsukamoto, K. A point mutation to the long terminal repeat of bovine leukemia virus related to viral productivity and transmissibility. Virology 2019, 537, 45–52.
Grez, M.; Zornig, M.; Nowock, J.; Ziegler, M. A single point mutation activates the Moloney murine leukemia virus long terminal repeat in embryonal stem cells. J. Virol. 1991, 65, 4691–4698.
Xu, X.; Kang, S.H.; Heidenreich, O.; Brown, D.A.; Nerenberg, M.I. Sequence requirements of ATF2 and CREB binding to the human T‐cell leukemia virus type 1 LTR R region. Virology 1996, 218, 362–371.
Pluta, A.; Rola‐Luszczak, M.; Douville, R.N.; Kuzmak, J. Bovine leukemia virus long terminal repeat variability: Identification of single nucleotide polymorphisms in regulatory sequences. Virol. J. 2018, 15, 165.
Zyrianova, I.M.; Kovalchuk, S.N. Bovine leukemia virus tax gene/Tax protein polymorphism and its relation to Enzootic Bovine Leukosis. Virulence 2020, 11, 80–87.
Derse, D. Bovine leukemia virus transcription is controlled by a virus‐encoded trans‐acting factor and by cis‐acting response elements. J. Virol. 1987, 61, 2462–2471.
Rose, N.J.; Richardson, J.H.; Desselberger, U.; Lever, A.M. Virus inactivation in a proportion of human T‐ cell leukaemia virus type I‐infected T‐cell clones arises through naturally occurring mutations. J. Gen. Virol. 2000, 81, 97–104.
Niewiesk, S.; Daenke, S.; Parker, C.E.; Taylor, G.; Weber, J.; Nightingale, S.; Bangham, C.R. Naturally occurring variants of human T‐cell leukemia virus type I Tax protein impair its recognition by cytotoxic T lymphocytes and the transactivation function of Tax. J. Virol. 1995, 69, 2649–2653.
Van Den Broeke, A.; Bagnis, C.; Ciesiolka, M.; Cleuter, Y.; Gelderblom, H.; Kerkhofs, P.; Griebel, P.; Mannoni, P.; Burny, A. In vivo rescue of a silent tax‐deficient bovine leukemia virus from a tumor‐derived ovine B‐cell line by recombination with a retrovirally transduced wild‐type tax gene. J. Virol. 1999, 73, 1054– 1065.
Durkin, K.; Rosewick, N.; Artesi, M.; Hahaut, V.; Griebel, P.; Arsic, N.; Burny, A.; Georges, M.; Van den Broeke, A. Characterization of novel Bovine Leukemia Virus (BLV) antisense transcripts by deep sequencing reveals constitutive expression in tumors and transcriptional interaction with viral microRNAs. Retrovirology 2016, 13, 33.
Willems, L.; Kettmann, R.; Dequiedt, F.; Portetelle, D.; Voneche, V.; Cornil, I.; Kerkhofs, P.; Burny, A.; Mammerickx, M. In vivo infection of sheep by bovine leukemia virus mutants. J. Virol. 1993, 67, 4078–4085.
Tajima, S.; Aida, Y. The region between amino acids 245 and 265 of the bovine leukemia virus (BLV) tax protein restricts transactivation not only via the BLV enhancer but also via other retrovirus enhancers. J. Virol. 2000, 74, 10939–10949.
Derse, D.; Caradonna, S.J.; Casey, J.W. Bovine leukemia virus long terminal repeat: A cell type‐specific promoter. Science 1985, 227, 317–320.
Willems, L.; Kettmann, R.; Burny, A. The amino acid (157‐197) peptide segment of bovine leukemia virus p34tax encompass a leucine‐rich globally neutral activation domain. Oncogene 1991, 6, 159–163.
Willems, L.; Grimonpont, C.; Heremans, H.; Rebeyrotte, N.; Chen, G.; Portetelle, D.; Burny, A.; Kettmann, R. Mutations in the bovine leukemia virus Tax protein can abrogate the long terminal repeat‐directed transactivating activity without concomitant loss of transforming potential. Proc. Natl. Acad. Sci. USA 1992, 89, 3957–3961.
Sakakibara, N.; Kabeya, H.; Ohashi, K.; Sugimoto, C.; Onuma, M. Epitope mapping of bovine leukemia virus transactivator protein Tax. J. Vet. Med. Sci. 1998, 60, 599–605.
Bai, L.; Takeshima, S.N.; Isogai, E.; Kohara, J.; Aida, Y. Novel CD8(+) cytotoxic T cell epitopes in bovine leukemia virus with cattle. Vaccine 2015, 33, 7194–7202.
Tajima, S.; Takahashi, M.; Takeshima, S.N.; Konnai, S.; Yin, S.A.; Watarai, S.; Tanaka, Y.; Onuma, M.; Okada, K.; Aida, Y. A mutant form of the tax protein of bovine leukemia virus (BLV), with enhanced transactivation activity, increases expression and propagation of BLV in vitro but not in vivo. J. Virol. 2003, 77, 1894–1903.
Terme, J.M.; Calvignac, S.; Duc Dodon, M.; Gazzolo, L.; Jordan, A. E box motifs as mediators of proviral latency of human retroviruses. Retrovirology 2009, 6, 81.
Calomme, C.; Dekoninck, A.; Nizet, S.; Adam, E.; Nguyen, T.L.; Van Den Broeke, A.; Willems, L.; Kettmann, R.; Burny, A.; Van Lint, C. Overlapping CRE and E box motifs in the enhancer sequences of the bovine leukemia virus 5′ long terminal repeat are critical for basal and acetylation‐dependent transcriptional activity of the viral promoter: Implications for viral latency. J. Virol. 2004, 78, 13848–13864.
Brooks, P.A.; Cockerell, G.L.; Nyborg, J.K. Activation of BLV transcription by NF‐kappa B and Tax. Virology 1998, 243, 94–98.
Duckett, C.S.; Perkins, N.D.; Kowalik, T.F.; Schmid, R.M.; Huang, E.S.; Baldwin, A.S., Jr.; Nabel, G.J. Dimerization of NF‐KB2 with RelA(p65) regulates DNA binding, transcriptional activation, and inhibition by an I kappa B‐alpha (MAD‐3). Mol. Cell Biol. 1993, 13, 1315–1322.
Bossone, S.A.; Asselin, C.; Patel, A.J.; Marcu, K.B. MAZ, a zinc finger protein, binds to c‐MYC and C2 gene sequences regulating transcriptional initiation and termination. Proc. Natl. Acad. Sci. USA 1992, 89, 7452– 7456.
Maity, G.; Haque, I.; Ghosh, A.; Dhar, G.; Gupta, V.; Sarkar, S.; Azeem, I.; McGregor, D.; Choudhary, A.; Campbell, D.R.; et al. The MAZ transcription factor is a downstream target of the oncoprotein Cyr61/CCN1 and promotes pancreatic cancer cell invasion via CRAF‐ERK signaling. J. Biol. Chem. 2018, 293, 4334–4349.
Kaplan, J.; Calame, K. The ZiN/POZ domain of ZF5 is required for both transcriptional activation and repression. Nucleic Acids Res. 1997, 25, 1108–1116.
Xiao, J.; Buehring, G.C. In vivo protein binding and functional analysis of cis‐acting elements in the U3 region of the bovine leukemia virus long terminal repeat. J. Virol. 1998, 72, 5994–6003.
Sagata, N.; Yasunaga, T.; Tsuzuku‐Kawamura, J.; Ohishi, K.; Ogawa, Y.; Ikawa, Y. Complete nucleotide sequence of the genome of bovine leukemia virus: Its evolutionary relationship to other retroviruses. Proc. Natl. Acad. Sci. USA 1985, 82, 677–681.
Inoue, E.; Matsumura, K.; Soma, N.; Hirasawa, S.; Wakimoto, M.; Arakaki, Y.; Yoshida, T.; Osawa, Y.; Okazaki, K. L233P mutation of the Tax protein strongly correlated with leukemogenicity of bovine leukemia virus. Vet. Microbiol. 2013, 167, 364–371.
Martinez, M.P.; Al‐Saleem, J.; Green, P.L. Comparative virology of HTLV‐1 and HTLV‐2. Retrovirology 2019, 16, 21.
Neto, W.K.; Da‐Costa, A.C.; de Oliveira, A.C.; Martinez, V.P.; Nukui, Y.; Sabino, E.C.; Sanabani, S.S. Correlation between LTR point mutations and proviral load levels among human T cell lymphotropic virus type 1 (HTLV‐1) asymptomatic carriers. Virol. J. 2011, 8, 535.
Merimi, M.; Klener, P.; Szynal, M.; Cleuter, Y.; Kerkhofs, P.; Burny, A.; Martiat, P.; Van den Broeke, A. Suppression of viral gene expression in bovine leukemia virus‐associated B‐cell malignancy: Interplay of epigenetic modifications leading to chromatin with a repressive histone code. J. Virol. 2007, 81, 5929–5939.
Achachi, A.; Florins, A.; Gillet, N.; Debacq, C.; Urbain, P.; Foutsop, G.M.; Vandermeers, F.; Jasik, A.; Reichert, M.; Kerkhofs, P.; et al. Valproate activates bovine leukemia virus gene expression, triggers apoptosis, and induces leukemia/lymphoma regression in vivo. Proc. Natl. Acad. Sci. USA 2005, 102, 10309– 10314.
Gillet, N.A.; Gutierrez, G.; Rodriguez, S.M.; de Brogniez, A.; Renotte, N.; Alvarez, I.; Trono, K.; Willems, L. Massive depletion of bovine leukemia virus proviral clones located in genomic transcriptionally active sites during primary infection. PLoS Pathog. 2013, 9, e1003687.
Murakami, H.; Yamada, T.; Suzuki, M.; Nakahara, Y.; Suzuki, K.; Sentsui, H. Bovine leukemia virus integration site selection in cattle that develop leukemia. Virus Res. 2011, 156, 107–112.
Lemasson, I.; Polakowski, N.J.; Laybourn, P.J.; Nyborg, J.K. Transcription factor binding and histone modifications on the integrated proviral promoter in human T‐cell leukemia virus‐I‐infected T‐cells. J. Biol. Chem. 2002, 277, 49459–49465.
Easley, R.; Carpio, L.; Guendel, I.; Klase, Z.; Choi, S.; Kehn‐Hall, K.; Brady, J.N.; Kashanchi, F. Human T‐ lymphotropic virus type 1 transcription and chromatin‐remodeling complexes. J. Virol. 2010, 84, 4755–4768.
Li, B.; Carey, M.; Workman, J.L. The role of chromatin during transcription. Cell 2007, 128, 707–719.
Murakami, H.; Uchiyama, J.; Suzuki, C.; Nikaido, S.; Shibuya, K.; Sato, R.; Maeda, Y.; Tomioka, M.; Takeshima, S.N.; Kato, H.; et al. Variations in the viral genome and biological properties of bovine leukemia virus wild‐type strains. Virus Res. 2018, 253, 103–111.
Debacq, C.; Sanchez Alcaraz, M.T.; Mortreux, F.; Kerkhofs, P.; Kettmann, R.; Willems, L. Reduced proviral loads during primo‐infection of sheep by Bovine Leukemia virus attenuated mutants. Retrovirology 2004, 1, 31.
Huelsenbeck, J.P.; Ronquist, F. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 2001, 17, 754–755.
Wingender, E.; Karas, H.; Knuppel, R. TRANSFAC database as a bridge between sequence data libraries and biological function. Pac. Symp. Biocomput. 1997, 2, 477‐485.
Van den Broeke, A.; Cleuter, Y.; Chen, G.; Portetelle, D.; Mammerickx, M.; Zagury, D.; Fouchard, M.; Coulombel, L.; Kettmann, R.; Burny, A. Even transcriptionally competent proviruses are silent in bovine leukemia virus‐induced sheep tumor cells. Proc. Natl. Acad. Sci. USA 1988, 85, 9263–9267.
Kerkhofs, P.; Heremans, H.; Burny, A.; Kettmann, R.; Willems, L. In vitro and in vivo oncogenic potential of bovine leukemia virus G4 protein. J. Virol. 1998, 72, 2554–2559.
Florins, A.; Gillet, N.; Boxus, M.; Kerkhofs, P.; Kettmann, R.; Willems, L. Even attenuated bovine leukemia virus proviruses can be pathogenic in sheep. J. Virol. 2007, 81, 10195–10200.
Rola‐Luszczak, M.; Finnegan, C.; Olech, M.; Choudhury, B.; Kuzmak, J. Development of an improved real time PCR for the detection of bovine leukaemia provirus nucleic acid and its use in the clarification of inconclusive serological test results. J. Virol. Methods 2013, 189, 258–264.
