Retrovirology | |
TRIM5α associates with proteasomal subunits in cells while in complex with HIV-1 virions | |
Edward M Campbell3  Jeremy Luban1  Seth L Robia2  Jaya Sastri3  Justin Rucci3  Sarah Sebastian1  Stéphane Hausmann1  Zana Lukic3  | |
[1] Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland;Department of Cell and Molecular Physiology, Stritch School of Medicine, Loyola University Chicago, Maywood IL, USA;Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood IL, USA | |
关键词: immunofluorescence; cytoplasmic bodies; proteasomal subunits; HIV-1; TRIM5α; | |
Others : 1209385 DOI : 10.1186/1742-4690-8-93 |
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received in 2011-08-31, accepted in 2011-11-12, 发布年份 2011 | |
【 摘 要 】
Background
The TRIM5 proteins are cellular restriction factors that prevent retroviral infection in a species-specific manner. Multiple experiments indicate that restriction activity requires accessory host factors, including E2-enzymes. To better understand the mechanism of restriction, we conducted yeast-two hybrid screens to identify proteins that bind to two TRIM5 orthologues.
Results
The only cDNAs that scored on repeat testing with both TRIM5 orthologues were the proteasome subunit PSMC2 and ubiquitin. Using co-immunoprecipitation assays, we demonstrated an interaction between TRIM5α and PSMC2, as well as numerous other proteasome subunits. Fluorescence microscopy revealed co-localization of proteasomes and TRIM5α cytoplasmic bodies. Forster resonance energy transfer (FRET) analysis indicated that the interaction between TRIM5 and PSMC2 was direct. Previous imaging experiments demonstrated that, when cells are challenged with fluorescently-labeled HIV-1 virions, restrictive TRIM5α orthologues assemble cytoplasmic bodies around incoming virion particles. Following virus challenge, we observed localization of proteasome subunits to rhTRIM5α cytoplasmic bodies that contained fluorescently labeled HIV-1 virions.
Conclusions
Taken together, the results presented here suggest that localization of the proteasome to TRIM5α cytoplasmic bodies makes an important contribution to TRIM5α-mediated restriction.
【 授权许可】
2011 Lukic et al; licensee BioMed Central Ltd.
【 预 览 】
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20150602101102768.pdf | 3632KB | download | |
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Figure 2. | 63KB | Image | download |
Figure 1. | 40KB | Image | download |
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【 参考文献 】
- [1]Lienlaf M, Hayashi F, Di Nunzio F, Tochio N, Kigawa T, Yokoyama S, Diaz-Griffero F: Contribution of E3-Ubiquitin Ligase Activity to HIV-1 Restriction by TRIM5{alpha}rh: Structure of the RING Domain of TRIM5{alpha}. J Virol 2011, 85:8725-8737.
- [2]Stremlau M, Owens CM, Perron MJ, Kiessling M, Autissier P, Sodroski J: The cytoplasmic body component TRIM5alpha restricts HIV-1 infection in Old World monkeys. Nature 2004, 427:848-853.
- [3]Reymond A, Meroni G, Fantozzi A, Merla G, Cairo S, Luzi L, Riganelli D, Zanaria E, Messali S, Cainarca S, et al.: The tripartite motif family identifies cell compartments. Embo J 2001, 20:2140-2151.
- [4]Sayah DM, Sokolskaja E, Berthoux L, Luban J: Cyclophilin A retrotransposition into TRIM5 explains owl monkey resistance to HIV-1. Nature 2004, 430:569-573.
- [5]Nisole S, Lynch C, Stoye JP, Yap MW: A Trim5-cyclophilin A fusion protein found in owl monkey kidney cells can restrict HIV-1. Proc Natl Acad Sci USA 2004, 101:13324-13328.
- [6]Sebastian S, Luban J: TRIM5alpha selectively binds a restriction-sensitive retroviral capsid. Retrovirology 2005, 2:40. BioMed Central Full Text
- [7]Stremlau M, Perron M, Lee M, Li Y, Song B, Javanbakht H, Diaz-Griffero F, Anderson DJ, Sundquist WI, Sodroski J: Specific recognition and accelerated uncoating of retroviral capsids by the TRIM5{alpha} restriction factor. Proc Natl Acad Sci USA 2006.
- [8]Wu X, Anderson JL, Campbell EM, Joseph AM, Hope TJ: Proteasome inhibitors uncouple rhesus TRIM5alpha restriction of HIV-1 reverse transcription and infection. Proc Natl Acad Sci USA 2006, 103:7465-7470.
- [9]Anderson JL, Campbell EM, Wu X, Vandegraaff N, Engelman A, Hope TJ: Proteasome inhibition reveals that a functional preintegration complex intermediate can be generated during restriction by diverse TRIM5 proteins. J Virol 2006, 80:9754-9760.
- [10]Diaz-Griffero F, Kar A, Perron M, Xiang SH, Javanbakht H, Li X, Sodroski J: Modulation of Retroviral Restriction and Proteasome Inhibitor-resistant Turnover by Changes in the TRIM5{alpha} B-box 2 Domain. J Virol 2007, 81:10362-10378.
- [11]Campbell EM, Perez O, Anderson JL, Hope TJ: Visualization of a proteasome-independent intermediate during restriction of HIV-1 by rhesus TRIM5alpha. J Cell Biol 2008, 180:549-561.
- [12]Rold CJ, Aiken C: Proteasomal degradation of TRIM5alpha during retrovirus restriction. PLoS Pathog 2008, 4:e1000074.
- [13]Pertel T, Hausmann S, Morger D, Zuger S, Guerra J, Lascano J, Reinhard C, Santoni FA, Uchil PD, Chatel L, et al.: TRIM5 is an innate immune sensor for the retrovirus capsid lattice. Nature 2011, 472:361-365.
- [14]Murata S, Yashiroda H, Tanaka K: Molecular mechanisms of proteasome assembly. Nat Rev Mol Cell Biol 2009, 10:104-115.
- [15]Geetha T, Seibenhener ML, Chen L, Madura K, Wooten MW: p62 serves as a shuttling factor for TrkA interaction with the proteasome. Biochem Biophys Res Commun 2008, 374:33-37.
- [16]Murata S: Multiple chaperone-assisted formation of mammalian 20S proteasomes. IUBMB Life 2006, 58:344-348.
- [17]Thylur RP, Senthivinayagam S, Campbell EM, Rangasamy V, Thorenoor N, Sondarva G, Mehrotra S, Mishra P, Zook E, Le PT, et al.: Mixed Lineage Kinase 3 modulates {beta}-catenin signaling in cancer cells. J Biol Chem 2011.
- [18]Campbell EM, Dodding MP, Yap MW, Wu X, Gallois-Montbrun S, Malim MH, Stoye JP, Hope TJ: TRIM5 alpha cytoplasmic bodies are highly dynamic structures. Mol Biol Cell 2007, 18:2102-2111.
- [19]Wojcik C, DeMartino GN: Intracellular localization of proteasomes. Int J Biochem Cell Biol 2003, 35:579-589.
- [20]Klare N, Seeger M, Janek K, Jungblut PR, Dahlmann B: Intermediate-type 20 S proteasomes in HeLa cells: "asymmetric" subunit composition, diversity and adaptation. J Mol Biol 2007, 373:1-10.
- [21]Dahlmann B, Ruppert T, Kuehn L, Merforth S, Kloetzel PM: Different proteasome subtypes in a single tissue exhibit different enzymatic properties. J Mol Biol 2000, 303:643-653.
- [22]Luciani A, Villella VR, Esposito S, Brunetti-Pierri N, Medina D, Settembre C, Gavina M, Pulze L, Giardino I, Pettoello-Mantovani M, et al.: Defective CFTR induces aggresome formation and lung inflammation in cystic fibrosis through ROS-mediated autophagy inhibition. Nat Cell Biol 2010, 12:863-875.
- [23]Hou Z, Kelly EM, Robia SL: Phosphomimetic mutations increase phospholamban oligomerization and alter the structure of its regulatory complex. J Biol Chem 2008, 283:28996-29003.
- [24]Kelly EM, Hou Z, Bossuyt J, Bers DM, Robia SL: Phospholamban oligomerization, quaternary structure, and sarco(endo)plasmic reticulum calcium ATPase binding measured by fluorescence resonance energy transfer in living cells. J Biol Chem 2008, 283:12202-12211.
- [25]O'Connor C, Pertel T, Gray S, Robia SL, Bakowska JC, Luban J, Campbell EM: p62/sequestosome-1 associates with and sustains the expression of retroviral restriction factor TRIM5alpha. J Virol 2010, 84:5997-6006.
- [26]Takalkar AM, Klibanov AL, Rychak JJ, Lindner JR, Ley K: Binding and detachment dynamics of microbubbles targeted to P-selectin under controlled shear flow. J Control Release 2004, 96:473-482.
- [27]Kenworthy AK: Imaging protein-protein interactions using fluorescence resonance energy transfer microscopy. Methods 2001, 24:289-296.
- [28]Ganser-Pornillos B, Chandrasekaran V, Pornillos O, Sodroski J, Sundquist WI, Yeager M: Hexagonal assembly of a restricting TRIM5alpha protein. Proc Natl Acad Sci USA 2010, in press.
- [29]Ganser-Pornillos BK, Chandrasekaran V, Pornillos O, Sodroski JG, Sundquist WI, Yeager M: Hexagonal assembly of a restricting TRIM5alpha protein. Proc Natl Acad Sci USA 2011, 108:534-539.
- [30]McDonald D, Vodicka MA, Lucero G, Svitkina TM, Borisy GG, Emerman M, Hope TJ: Visualization of the intracellular behavior of HIV in living cells. J Cell Biol 2002, 159:441-452.
- [31]Diaz-Griffero F, Li X, Javanbakht H, Song B, Welikala S, Stremlau M, Sodroski J: Rapid turnover and polyubiquitylation of the retroviral restriction factor TRIM5. Virology 2006, 349:300-315.
- [32]Yamauchi K, Wada K, Tanji K, Tanaka M, Kamitani T: Ubiquitination of E3 ubiquitin ligase TRIM5 alpha and its potential role. FEBS J 2008, 275:1540-1555.
- [33]Rosenqvist E, Jossang T, Feder J: Thermal properties of human IgG. Mol Immunol 1987, 24:495-501.
- [34]Terry BR, Matthews EK, Haseloff J: Molecular characterisation of recombinant green fluorescent protein by fluorescence correlation microscopy. Biochem Biophys Res Commun 1995, 217:21-27.
- [35]Perez-Caballero D, Hatziioannou T, Zhang F, Cowan S, Bieniasz PD: Restriction of Human Immunodeficiency Virus Type 1 by TRIM-CypA Occurs with Rapid Kinetics and Independently of Cytoplasmic Bodies, Ubiquitin, and Proteasome Activity. J Virol 2005, 79:15567-15572.
- [36]Song B, Diaz-Griffero F, Park DH, Rogers T, Stremlau M, Sodroski J: TRIM5alpha association with cytoplasmic bodies is not required for antiretroviral activity. Virology 2005, 343:201-211.
- [37]Gyuris J, Golemis E, Chertkov H, Brent R: Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell 1993, 75:791-803.
- [38]Sastri J, O'Connor C, C D, M M, P P, Diaz-Griffero F, Campbell EM: Identification of residues within the L2 region of TRIM5α that are required for retroviral restriction and cytoplasmic body localization. Virology 2010, 405:259-266.
- [39]O'Connor C, Pertel T, Gray S, Robia SL, Bakowska JC, Luban J, Campbell EM: p62/Sequestosome1 associates with and sustains the expression of the retroviral restriction factor TRIM5{alpha}. J Virol 2010.
- [40]Hirano Y, Hayashi H, Iemura S, Hendil KB, Niwa S, Kishimoto T, Kasahara M, Natsume T, Tanaka K, Murata S: Cooperation of multiple chaperones required for the assembly of mammalian 20S proteasomes. Mol Cell 2006, 24:977-984.
- [41]Durocher Y, Perret S, Kamen A: High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res 2002, 30:E9.
- [42]Campbell EM, Nunez R, Hope TJ: Disruption of the actin cytoskeleton can complement the ability of Nef to enhance human immunodeficiency virus type 1 infectivity. J Virol 2004, 78:5745-5755.
- [43]Sastri J, O'Connor C, Danielson CM, McRaven M, Perez P, Diaz-Griffero F, Campbell EM: Identification of residues within the L2 region of rhesus TRIM5alpha that are required for retroviral restriction and cytoplasmic body localization. Virology 2010, 405:259-266.
- [44]Zal T, Gascoigne NR: Photobleaching-corrected FRET efficiency imaging of live cells. Biophys J 2004, 86:3923-3939.