The Molecular Virology Laboratory primarily conducts research into the processing of viral messenger RNA (mRNA), with a specialized focus on the human immunodeficiency virus type 1 (HIV-1). The HIV-1 mRNA processing steps include splicing, polyadenylation, transport, cellular localisation, degradation and translation of mRNA into protein. Each step provides a potential means to modulate viral replication, infectivity and pathogenesis, therefore presenting likely targets for antiviral drug design. We study both the HIV-1 encoded and the host cell-derived control mechanisms that determine the efficiency of viral protein expression. RNA-dependent mechanisms, such as RNA interference (RNAi) support cellular antiviral defence pathways that are powerful innate mechanisms that restrict viral replication. We are investigating the convergence of these innate antiviral pathways with adaptive immunity.
From the perspective of virus, our studies of HIV-1 have shown that splicing generates a large array of alternatively spliced mRNAs from the nascent HIV-1 unspliced precursor-mRNA. Many of these mRNAs differ only in their 5' untranslated regions (UTR) and encode the same HIV-1 proteins. We have identified that env mRNAs use a ribosome shunting mechanism during translation, and our studies have identified mRNA elements that control this mechanism. Other aspects that determine translation control, such as the role of viral non-coding introns during viral replication, are also under investigation in our laboratory.
From the cellular aspect, we are focusing on the mechanisms that silence the expression of viral proteins in the astrocytes of brain. Astrocytes are important natural target cells for HIV-1 and their unusual refractory infection serves as a model that may yield novel approaches to restrict HIV-1 in other infected cell populations. Finally, we are applying our understanding of these fine details of HIV-1 and simian immunodeficiency virus (SIV) gene expression to the preparation of novel HIV-1 vaccines.
- RNA-mediated control of HIV latency
- Mechanisms controlling translation of HIV-1 envelope protein
- Modulation of HIV gene expression by RNA binding proteins
- Modulation of HIV-1 RNA translation
- Regulation of mRNA splicing and stability in macrophages
- Control of HIV gene expression by siRNA
- Replicon based HIV vaccine development
- Vaccine for HIV neutralising antibodies
- Assays for HIV neutralising antibodies
- Immunomodulatory activities of HIV-1 Vpu protein
Head and Associate Professor
Assoc Prof Damian Purcell: see profile
Dr Rob Center (SRO)
Dr Con Sonza (RF)
Jane Howard (RA)
Delphine Allen (RA)
Charlene Mackenzie (RA)
Daniel Yuen (PhD student)
Jonathan Jacobson (PhD student)
Talia Mota (PhD student)
Giovana Bernadi (PhD student)
Behnaz Heydarchi (PhD student)
Joanna Yeo (Hons student)
Chris Gonelli (Hons student)
- Wheatley, AK, Kramski M, Alexander MR, Toe JG, Center RJ, Purcell DF. Co-Expression of miRNA Targeting the Expression of PERK, but Not PKR, Enhances Cellular Immunity from an HIV-1 Env DNA Vaccine. PLoS ONE 2011; 6: e18225.
- Saleh S, Wightman F, Ramayanake S, Alexander M, Kumar N, da Fonseca Pereira C, Purcell DFJ, Cameron PU , Lewin SR. HIV expression and reactivation in a chemokine induced model of latent HIV infection of resting CD4+ T cells. Retrovirology 2011; 8: 80
- Suzuki K, Ishida T, Yamagishi M, Ahlenstiel C, Swaminathan S, Marks K, Murray D, McCartney EM, Beard MR, Alexander M, Purcell DFJ, Cooper DA, Watanabe T, Kelleher AD. Transcriptional gene silencing of HIV-1 through promoter targeted RNA is highly specific. RNA Biology 2011; 8: 1
- Fernandez S, Tanaskovic S, Helbig K, Rajasuriar R, Kramski M, Murray JM, Beard M, Purcell D, Lewin SR, Price P, French MA. CD4+ T-cell deficiency in HIV patients responding to ART is associated with increased expression of interferon-stimulated genes in CD4+ T-cells. J Infect Diseases 2011; 204: 1927-1935.
- Lim, A, Amini A, D'Orsogna LJ, Rajasuriar R, Kramski M, Lewin SR, Purcell DF, Price P, French MA, Antibody and B-cell responses may control circulating lipopolysaccharide in patients with HIV infection. Aids 2011; 25: 1379-1383.
- Kramski M, Gaeguta AJ, Lichtfuss GF, Rajasuriar R, Crowe SM, French MA, Lewin SR, Center RJ, Purcell DFJ. Novel Sensitive Real-Time PCR for Quantification of Bacterial 16S rRNA Genes in Plasma of HIV-Infected Patients as a Marker for Microbial Translocation. Journal of Clinical Microbiology 2011; 49: 3691-3693.
- Kirkegaard T, Wheatley A, Melchjorsen J, Bahrami S, Pedersen FS, Center RJ, Purcell DFJ, Ostergaard L, Duch M, Tolstrup M. Induction of humoral and cellular immune responses against the HIV-1 envelope protein using gamma-retroviral virus-like particles. Virology Journal 2011; 8: 381.
- Byakwaga H, Kelly M, Purcell DFJ, French MA, Amin J, Lewin SR, Haskelberg H, Kelleher AD, Garsia R, Boyd MA, Cooper DA, Emery S, Grp CS. Intensification of Antiretroviral Therapy With Raltegravir or Addition of Hyperimmune Bovine Colostrum in HIV-Infected Patients With Suboptimal CD4(+) T-Cell Response: A Randomized Controlled Trial. Journal of Infectious Diseases 2011; 204; 1532-1540.
- Alexander MR, Wheatley AK, Center RJ, Purcell D F. Efficient transcription through an intron requires the binding of an Sm-type U1 snRNP with intact stem loop II to the splice donor. Nucl Acids Res 2010; 38: 3041-3053.
- Wade J, Sterjovski J, Gray L, Roche M, Chiavaroli L, Ellett A, Jakobsen MR, Cowley D, Pereira Cda F, Saksena N, Wang B, Purcell DF, Karlsson I, Fenyo EM, Churchill M, Gorry PR. Enhanced CD4+ cellular apoptosis by CCR5-restricted HIV-1 envelope glycoprotein variants from patients with progressive HIV-1 infection. Virology 2010; 396: 246-255.
- Wilson R, Purcell D, Netter HJ, Revill PA. Does RNA interference provide new hope for control of chronic hepatitis B infection? Antivir Ther 2009; 147; 879-889.
- Center RJ, Wheatley AK, Campbell SM, Gaeguta AJ, Peut V, Alcantara S, Siebentritt C, Kent SJ, Purcell DFJ. Induction of HIV-1 subtype B and AE-specific neutralizing antibodies in mice and macaques with DNA prime and recombinant gp140 protein boost regimens. Vaccine 2009; 27: 6605-12.
- Peut V, Campbell S, Gaeguta A, Center RJ, Wilson K, Alcantara S, Fernandez CS, Purcell DF, Kent SJ. Balancing reversion of CTL and neutralizing antibody escape mutations within HIV-1 Env upon transmission. J Virol 2009; 83: 8986-92.
- Gray L, Roche M, Churchill MJ, Sterjovski J, Ellett A, Poumbourios P, Sheffief S, Wang B, Saksena N, Purcell DF, Wesselingh S, Cunningham AL, Brew BJ, Gabuzda D, Gorry PR. Tissue-specific sequence alterations in the human immunodeficiency virus type 1 envelope favoring CCR5 usage contribute to persistence of dual-tropic virus in the brain. J Virol 2009; 83: 5430-41.
- Daniels SM, Melendez-Peña CE, Scarborough RJ, Daher A, Christensen HS, El Far M, Purcell DF, Lainé S, Gatignol A. Characterization of the TRBP domain required for Dicer interaction and function in RNA interference. BMC Mol BiolM/ 2009; 10: 38.
- Loh L, Reece JC, Fernandez CS, Alcantara S, Center R, Howard J, Purcell DF, Balamurali M, Petravic J, Davenport MP, Kent SJ. . Complexity of the inoculum determines the rate of reversion of SIV Gag CD8 T cell mutant virus and outcome of infection. PLoS Pathog 2009; 5: e1000378.
- Kelleher AD, Purcell DF. Controlling HIV infection: HIV co-receptor gets the finger. Immunol Cell Biol 2008; 86: 641-2.
- Borggren M, Repits J, Kuylenstierna C, Sterjovski J, Churchill MJ, Purcell DF, Karlsson A, Albert J, Gorry PR, Jansson M. Evolution of DC-SIGN use revealed by fitness studies of R5 HIV-1 variants emerging during AIDS progression. Retrovirology 20085:28.
- Miller A, Center RJ, Stambas J, Deliyannis G, Doherty PC, Howard JL, Turner SJ, Purcell DF. Sindbis virus vectors elicit hemagglutinin-specific humoral and cellular immune responses and offer a dose-sparing strategy for vaccination. Vaccine 2008; 26: 5641-5648.
- Lewin SR, Murray JM, Solomon A, Wightman F, Cameron PU, Purcell DF, Zaunders JJ, Grey P, Bloch M, Smith D, Cooper DA, Kelleher AD. Virologic determinants of success after structured treatment interruptions of antiretrovirals in acute HIV-1 infection. J Acquir Immune Defic Syndr 2008; 47: 140-47.
- Dowling D, Nasr-Esfahani S, Tan CH, O'Brien K, Howard JL, Jans DA, Purcell DF, Stoltzfus CM, Sonza S. HIV-1 infection induces changes in expression of cellular splicing factors that regulate alternative viral splicing and virus production in macrophages. Retrovirology 2008; 5: 18
- Repits J, Sterjovski J, Badia-Martinez D, Mild M, Gray L, Churchill MJ, Purcell DF, Karlsson A, Albert J, Fenyo EM, Achour A, Gorry PR, Jansson M. Primary HIV-1 R5 isolates from end-stage disease display enhanced viral fitness in parallel with increased gp120 net charge. Virology 2008; 379 1: 125-34.
- Harrison DN, Gazina EV, Purcell DF, Anderson DA, Petrou S. Amiloride derivatives inhibit coxsackievirus B3 RNA replication. J Virol 2008; 82 3: 1465-73.
- Cameron PU, Handley AJ, Baylis DC, Solomon AE, Bernard N, Purcell DF, Lewin SR. Preferential infection of dendritic cells during human immunodeficiency virus type 1 infection of blood leukocytes. J Virol 2007; 81: 2297-306.
- Rollman E, Smith MZ, Brooks A, Purcell DFJ, Zuber B, Ramshaw IA, Kent SJ. Killing kinetics of SIV-specific CD8+ T cells: implications for HIV vaccine strategies. Journal of Immunology 2007; 179: 4571-4579.
- Anderson JL, Johnson AT, Howard JL, Purcell DF. Both linear and discontinuous ribosome scanning are used for translation initiation from bicistronic human immunodeficiency virus type 1 env mRNAs. J Virol 2007; 81: 4664-76.
- Krummheuer J, Johnson AT, Hauber I, Kammler S, Anderson JL, Hauber J, Purcel DFl*, Schaal H.* A minimal uORF within the HIV-1 vpu leader allows efficient translation initiation at the downstream env AUG. Virology 2007; 363: 261-71. (* Joint senior authors)
- Christensen HS, Daher A, Soye KJ, Frankel LB, Alexander MR,Laine S, Bannwarth S, Ong CL, Chung SW, Campbell SM, Purcell DF*, Gatignol A*. Small interfering RNAs against the TAR RNA binding protein, TRBP, a Dicer cofactor, inhibit human immunodeficiency virus type 1 long terminal repeat expression and viral production. J Virol 2007; 81: 5121-31 (* Joint senior authors)
- Sinck L, Richer D, Howard J, Alexander M, Purcell DF, Marquet R, Paillart JC. In vitro dimerization of human immunodeficiency virus type 1 (HIV-1) spliced RNAs. RNA 2007; 13: 2141-50.
- Sterjovski J, Churchill MJ, Ellett A, Gray LR, Roche MJ, Dunfee RL, Purcell DF, Saksena N, Wang B, Sonza S, Wesselingh SL, Karlsson I, Fenyo EM, Gabuzda D, Cunningham AL, Gorry PR. Asn 362 in gp120 contributes to enhanced fusogenicity by CCR5-restricted HIV-1 envelope glycoprotein variants from patients with AIDS. Retrovirology 2007; 4: 89.
- Gray, L, Churchill MJ, Keane N, Sterjovski J, Ellett AM, Purcell DF, Poumbourios, Kol C, Wang B, Saksena NK, Wesselingh SL, Price P, French M, Gabuzda D, Gorry PR. Genetic and Functional Analysis of R5X4 Human Immunodeficiency Virus Type 1 Envelope Glycoproteins Derived from Two Individuals Homozygous for the CCR5∆32 Allele. J Virol 2006; 80: 3684-91.
- Kelleher AD, Puls R, Bebbington M, Boyle D, Ffrench R, Kent S, Kippax S, Law M, Purcell DFJ, Ramshaw I, Cooper D, Emery S. A Randomised, Placebo-controlled Phase I Trial of DNA Prime, Recombinant Fowlpox Virus Boost Prophylactic Vaccine for HIV-1. Aids 2006; 20: 294-7.
- Ong CL, Thorpe JC, Gorry PR, Bannwarth S, Jawarowski A, Howard JL, Chung S, Mouland A, Gatignol A, Purcell DFJ. Low astrocyte TRBP levels support an innate HIV-1 resistance by enhancing the PKR antiviral response. J Virol 2005; 79: 12763–12772.
- Sonza S, H. Mutimer HP, O'Brien K, Ellery P, Howard JL, Axelrod JH, Deacon NJ, Crowe SM, Purcell DF. Selectively reduced tat mRNA heralds the decline in productive human immunodeficiency virus type 1 infection in monocyte-derived macrophages. J Virol 2002; 76: 12611-21.
- Sonza S, Mutimer H, Oelrichs R, Jardin D, Harvey K, Dunne A, Purcell DF, Birch C, Crowe S. Monocytes harbour replication-competent, non-latent HIV-1 in patients on highly active antiretroviral therapy. AIDS 2001; 15: 1-6. Kent SJ, Dale CJ, Preiss S, Mills J, Campagna D, Purcell DF. Vaccination with attenuated simian immunodeficiency virus by DNA inoculation. J Virol 2001; 75: 11930-4.
- Kent SJ, Cameron PU, Reece JC, Thompson PR, Purcell DF. Attenuated and wild-type HIV-1 infections and long terminal repeat- mediated gene expression from plasmids delivered by gene gun to human skin ex vivo and macaques in vivo. Virology 2001; 287: 71-8.
- Gorry PR, Howard JL, Churchill MJ, Anderson JL, Cunningham A, Adrian D, McPhee DA, Purcell DFJ. Diminished production of Human Immunodeficiency Virus Type 1 in astrocytes results from inefficient translation of gag, env, and nef mRNAs despite efficient expression of Tat and Rev. J Virol 1999; 73: 353-361.
- Ristevski S, Purcell DFJ, Marshall J, Campagna D, Nouri S, Fenton SP, McPhee DA, Kannourakis G. Novel type-D endogenous retroviral particles expressed at high levels in a SCID mouse thymic lymphoma. J Virol 1999; 73: 4662-4669.
- Gorry P, Purcell D, Howard J, McPhee D. Restricted HIV-1 infection of human astrocytes: potential role of nef in the regulation of virus replication. J NeuroVirol 1998; 4: 377-386.
- Purcell DFJ, Broscius CM, Vanin EF, Buckler CE, Nienhuis AW, Martin MA. An array of MuLV-related elements are transmitted and expressed in a primate recipient of retroviral gene transfer. J Virol 1996; 70: 887-897. (cited 57 times)
- Purcell DFJ, Martin MA. Alternative splicing of human immunodeficiency virus type 1 mRNA modulates viral protein expression, replication, and infectivity. J Virol 1993; 67: 6365-6378.