Infection by agnoprotein-negative mutants of polyomavirus JC and SV40 results in the release of virions that are mostly deficient in DNA content

Virology Journal

Mahmut Safak¹  Martyn K White¹  Abdullah S Saribas¹  Ilker K Sariyer¹ 

[1] Department of Neuroscience, Laboratory of Molecular Neurovirology, Temple University School of Medicine, 3500 N. Broad Street, Philadelphia PA 19140 USA

关键词: virion release;    transcription;    replication;    SV40;    BK virus;    JC virus;   

Others  :  1156636
DOI  :  10.1186/1743-422X-8-255

 received in 2011-01-19, accepted in 2011-05-24,  发布年份 2011

【 摘 要 】

Background

Human polyomavirus JC (JCV) is the etiologic agent of a brain disease, known as progressive multifocal leukoencephalopathy (PML). The JCV genome encodes a small multifunctional phospho-protein, agnoprotein, from the late coding region of the virus, whose regulatory functions in viral replication cycle remain elusive. In this work, the functional role of JCV and SV40 agnoproteins in virion release was investigated using a point mutant (Pt) of each virus, where the ATG codon of agnoprotein was mutated to abrogate its expression.

Results

Analysis of both viral protein expression and replication using Pt mutant of each virus revealed that both processes were substantially down-regulated in the absence of agnoprotein compared to wild-type (WT) virus. Complementation studies in cells, which are constitutively expressing JCV agnoprotein and transfected with the JCV Pt mutant genome, showed an elevation in the level of viral DNA replication near to that observed for WT. Constitutive expression of large T antigen was found to be not sufficient to compensate the loss of agnoprotein for efficient replication of neither JCV nor SV40 in vivo. Examination of the viral release process for both JCV and SV40 Pt mutants showed that viral particles are efficiently released from the infected cells in the absence of agnoprotein but were found to be mostly deficient in viral DNA content.

Conclusions

The results of this study provide evidence that agnoprotein plays an important role in the polyomavirus JC and SV40 life cycle. Infection by agnoprotein-negative mutants of both viruses results in the release of virions that are mostly deficient in DNA content.

【 授权许可】

   
2011 Sariyer et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Collette Y, Olive D: The primate lentivirus-encoded Nef protein can regulate several steps of the viral replication cycle. Virology 1999, 265:173-177.
• [2]Levy ND, Refaeli Y, Weiner DB: Extracellular Vpr protein increases cellular permissiveness to human immunodeficiency virus replication and reactivates virus from latency. J Virol 1995, 68:1243-1252.
• [3]Spina CA, Kwoh TJ, Chowers MY, Guatelli JC, Richman DD: The importance of nef in the induction of human immunodeficiency virus type 1 replication from primary quiescent CD4 lymphocytes. J Exp Med 1994, 179:115-123.
• [4]Cohen EA, Subbramanian RA, Gottlinger HG: Role of auxiliary proteins in retroviral morphogenesis. Cur Top Microbiol Immonol 1996, 214:219-235.
• [5]Cullen B: HIV-1 auxiliary proteins:making connections in dying cell. Cell 1998, 93:685-692.
• [6]Diaz-Guerra M, Esteban M: Vaccinia virus nucleoside triphosphate phosphorylase I controls early and late gene expression by regulating the rate of transcription. J Virol 1993, 67:7561-7572.
• [7]Frisque RJ, White FA: The molecular biology of JC virus, causative agent of progressive multifocal leukoencephalopathy. Edited by Totowa, NJ. Humana Press Inc.; 1992.
• [8]Perini G, Wagner S, Green MR: Recognition to bZIP proteins by the human T-cell leukaemia virus transactivator Tax. Nature 1995, 376:602-605.
• [9]Wei P, Garber ME, Fang S-M, Fisher WH, Jones KA: A novel CDK-9 associated C-type cyclin interacts directly with HIV-1 tat and mediates its high-affinity, loop-specific binding to TAR RNA. Cell 1998, 92:451-462.
• [10]Safak M, Barrucco R, Darbinyan A, Okada Y, Nagashima K, Khalili K: Interaction of JC virus agno protein with T antigen modulates transcription and replication of the viral genome in glial cells. J Virol 2001, 75:1476-1486.
• [11]Garcia-Sastre A, Egorov A, Matassov D, Brant S, Levy DE, Dublin JE, Palase P, Master T: Influenza virus lacking the NS1 gene replicates in interferon-deficient systems. Virology 1998, 252:324-330.
• [12]Fisher U, Huber J, Boelens WC, Mattaj IW, Baltimore D: The HIV-1 rev activation domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs. Cell 1995, 82:475-483.
• [13]Michaud G, Zachary A, Rao VB, Black LW: Membrane associated assembly of a phage T4 DNA entrance vertex structure studied with expression vectors. J Mol Med 1989, 209:667-681.
• [14]Terwillinger EF, Cohen EA, Lu YC, Sodroski JG, Hasetine WA: Functional role of human immunodeficiency virus type 1 vpu. Proc Natl Acad Sci USA 1989, 86:5163-5167.
• [15]Strebel K, Klimkait T, Maldarelli F, Martin MA: Molecular and biochemical analyses of human immunodeficiency virus type 1 vpu protein. J Virol 1989, 63:3784-3791.
• [16]Darbinyan A, Darbinian N, Safak M, Radhakrishnan S, Giordano A, Khalili K: Evidence for dysregulation of cell cycle by human polyomavirus, JCV, late auxiliary protein. Oncogene 2002, 21:5574-5581.
• [17]Gilbert J, Nomura S, Anderson CW, George K: Identification of the SV40 agnoproduct: a DNA binding protein. Nature 1981, 291:346-349.
• [18]Rinaldo CH, Traavik T, Hey A: The agnogene of the human polyomavirus BK is expressed. J Virol 1998, 72:6233-6236.
• [19]Safak M, Sadowska B, Barrucco R, Khalili K: Functional interaction between JC virus late regulatory agnoprotein and cellular Y-box binding transcription factor, YB-1. J Virol 2002, 76:3828-3838.
• [20]Okada Y, Sawa H, Endo S, Orba Y, Umemura T, Nishihara H, Stan AC, Tanaka S, Takahashi H, Nagashima K: Expression of JC virus agnoprotein in progressive multifocal leukoencephalopathy brain. Acta Neuropathol (Berl) 2002, 104:130-136.
• [21]Khalili K, White MK, Sawa H, Nagashima K, Safak M: The agnoprotein of polyomaviruses: A multifunctional auxiliary protein. J Cell Physiol 2005, 204:1-7.
• [22]Achim CL, Wiley CA: Expression of major histocompatibility complex antigens in the brains of patients with progressive multifocal leukoencephalopathy. J Neuropathol Exp Neurol 1992, 51:257-263.
• [23]Berger JR, Tornatore C, Major EO, Bruce J, Shapshak P, Yoshioka M, Houff S, Sheremata W, Horton GF, Landy H: Relapsing and remitting human immunodeficiency virus-associated leukoencephalomyelopathy. Ann Neurol 1992, 31:34-38.
• [24]Major EO, Amemiya K, Tornatore CS, Houff SA, Berger JR: Pathogenesis and molecular biology of progressive multifocal leukoencephalopathy, the JC virus-induced demyelinating disease of the human brain. Clin Microbiol Rev 1992, 5:49-73.
• [25]Berger JR, Concha M: Progressive multifocal leukoencephalopathy: the evolution of a disease once considered rare. J Neurovirol 1995, 1:5-18.
• [26]Sariyer IK, Khalili K, Safak M: Dephosphorylation of JC virus agnoprotein by protein phosphatase 2A: inhibition by small t antigen. Virology 2008, 375:464-479.
• [27]Hay N, Skolnik-David H, Aloni Y: Attenuation in the control of SV40 gene expression. Cell 1982, 29:183-193.
• [28]Haggerty S, Walker DL, Frisque RJ: JC virus-simian virus 40 genomes containing heterologous regulatory signals and chimeric early regions: identification of regions restricting transformation by JC virus. J Virol 1989, 63:2180-2190.
• [29]Alwine JC: Evidence for simian virus 40 late transcriptional control: mixed infections of wild-type simian virus 40 and a late leader deletion mutant exhibit trans effects on late viral RNA synthesis. J Virol 1982, 42:798-803.
• [30]Margolskee RF, Nathans D: Suppression of a VP1 mutant of simian virus 40 by missense mutations in serine codons of the viral agnogene. J Virol 1983, 48:405-409.
• [31]Ng SC, Mertz JE, Sanden-Will S, Bina M: Simian virus 40 maturation in cells harboring mutants deleted in the agnogene. J Biol Chem 1985, 260:1127-1132.
• [32]Hou-Jong MH, Larsen SH, Roman A: Role of the agnoprotein in regulation of simian virus 40 replication and maturation pathways. J Virol 1987, 61:937-939.
• [33]Barkan A, Welch RC, Mertz JE: Missense mutations in the VP1 gene of simian virus 40 that compensate for defects caused by deletions in the viral agnogene. J Virol 1987, 61:3190-3198.
• [34]Resnick J, Shenk T: Simian virus 40 agnoprotein facilitates normal nuclear location of the major capsid polypeptide and cell-to-cell spread of virus. J Virol 1986, 60:1098-1106.
• [35]Nomura S, Khoury G, Jay G: Subcellular localization of the simian virus 40 agnoprotein. J Virol 1983, 45:428-433.
• [36]Sariyer IK, Akan I, Palermo V, Gordon J, Khalili K, Safak M: Phosphorylation mutants of JC virus agnoprotein are unable to sustain the viral infection cycle. J Virol 2006, 80:3893-3903.
• [37]Johannessen M, Myhre MR, Dragset M, Tummler C, Moens U: Phosphorylation of human polyomavirus BK agnoprotein at Ser-11 is mediated by PKC and has an important regulative function. Virology 2008, 379:97-109.
• [38]Myhre MR, Olsen GH, Gosert R, Hirsch HH, Rinaldo CH: Clinical polyomavirus BK variants with agnogene deletion are non-functional but rescued by trans-complementation. Virology 2010, 398:12-20.
• [39]Suzuki T, Orba Y, Okada Y, Sunden Y, Kimura T, Tanaka S, Nagashima K, Hall WW, Sawa H: The human polyoma JC virus agnoprotein acts as a viroporin. PLoS Pathog 2010, 6:e1000801.
• [40]Major EO, Miller AE, Mourrain P, Traub RG, de Widt E, Sever J: Establishment of a line of human fetal glial cells that supports JC virus multiplication. Proc Natl Acad Sci USA 1985, 82:1257-1261.
• [41]Hirt B: Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 1967, 26:365-369.
• [42]Ansari SA, Safak M, Del Valle L, Enam S, Amini S, Khalili K: Cell cycle regulation of NF-kappa b-binding activity in cells from human glioblastomas. Exp Cell Res 2001, 265:221-233.
• [43]Sadowska B, Barrucco R, Khalili K, Safak M: Regulation of human polyomavirus JC virus gene transcription by AP-1 in glial cells. J Virol 2003, 77:665-672.
• [44]Ziegler K, Bui T, Frisque RJ, Grandinetti A, Nerurkar VR: A rapid in vitro polyomavirus DNA replication assay. J Virol Methods 2004, 122:123-127.
• [45]Lynch KJ, Frisque RJ: Identification of critical elements within the JC virus DNA replication origin. J Virol 1990, 64:5812-5822.
• [46]Lynch KJ, Frisque RJ: Factors contributing to the restricted DNA replicating activity of JC virus. Virology 1991, 180:306-317.
• [47]Lynch KJ, Haggerty S, Frisque RJ: DNA replication of chimeric JC virus-simian virus 40 genomes. Virology 1994, 204:819-822.
• [48]Nesper J, Smith RW, Kautz AR, Sock E, Wegner M, Grummt F, Nasheuer HP: A cell-free replication system for human polyomavirus JC DNA. J Virol 1997, 71:7421-7428.
• [49]Sock E, Wegner M, Fortunato EA, Grummt F: Large T-antigen and sequences within the regulatory region of JC virus both contribute to the features of JC virus DNA replication. Virology 1993, 197:537-548.