Downregulation of APOBEC3G by xenotropic murine leukemia-virus related virus (XMRV) in prostate cancer cells
- Abhinav Dey†1,
- Chinmay Kumar Mantri†1,
- Jui Pandhare-Dash†1,
- Bindong Liu1,
- Siddharth Pratap2 and
- Chandravanu Dash1Email author
© Dey et al; licensee BioMed Central Ltd. 2011
Received: 26 September 2011
Accepted: 12 December 2011
Published: 12 December 2011
Xenotropic murine leukemia virus (MLV)-related virus (XMRV) is a gammaretrovirus that was discovered in prostate cancer tissues. Recently, it has been proposed that XMRV is a laboratory contaminant and may have originated via a rare recombination event. Host restriction factor APOBEC3G (A3G) has been reported to severely restrict XMRV replication in human peripheral blood mononuclear cells. Interestingly, XMRV infects and replicates efficiently in prostate cancer cells of epithelial origin. It has been proposed that due to lack off or very low levels of A3G protein XMRV is able to productively replicate in these cells.
This report builds on and challenges the published data on the absence of A3G protein in prostate epithelial cells lines. We demonstrate the presence of A3G in prostate epithelial cell lines (LNCaP and DU145) by western blot and mass spectrometry. We believe the discrepancy in A3G detection is may be due to selection and sensitivity of A3G antibodies employed in the prior studies. Our results also indicate that XMRV produced from A3G expressing LNCaP cells can infect and replicate in target cells. Most importantly our data reveal downregulation of A3G in XMRV infected LNCaP and DU145 cells.
We propose that XMRV replicates efficiently in prostate epithelial cells by downregulating A3G expression. Given that XMRV lacks accessory proteins such as HIV-1 Vif that are known to counteract A3G function in human cells, our data suggest a novel mechanism by which retroviruses can counteract the antiviral effects of A3G proteins.
KeywordsXMRV APOBEC3G Retrovirus Prostate
Xenotropic murine leukemia-virus related virus (XMRV) is a member of the gammaretrovirus family that was first detected in human prostate tumors . Although initial studies supported the presence of XMRV in prostate cancer tissues [2–4], since then several laboratories have failed to detect the virus in cohorts of prostate cancer patients [5–9]. Very recently, Paprotka et al. (2011) have challenged an association of XMRV with human diseases . These authors have reported that XMRV may have originated by a rare recombination event during tumor passaging in mice. Therefore, it has been proposed that XMRV is a laboratory contaminant and not a human pathogen. Nevertheless, being a newly discovered gammaretrovirus and having the ability to infect human cells, XMRV may serve as a model gammaretrovirus to further our understanding of retroviral biology.
Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3) proteins, APOBEC3A to APOBEC3G are a class of cytidine deaminases that has been reported to restrict retroviral replication in humans . In case of HIV-1, the restriction by APOBEC3G (A3G) and APOBEC3F (A3F) are counteracted by Vif that degrades A3 proteins via proteosomal degradation . It has been reported that XMRV replication can be inhibited by A3 proteins such as A3G, A3B, A3F, and murine APOBEC3 (mA3) [13–17]. Although hypermutation of XMRV genome in A3G/A3F-expressing peripheral blood mononuclear cells (PBMCs) severely restrict XMRV replication in human blood, infected PBMCs have been suggested to serve as source of infectious XMRV . Given that XMRV is a simple retrovirus and does not encode accessory proteins that are known to counteract A3G/A3F, these observations suggest XMRV cannot survive the restriction of innate immunity for productive infection in humans.
XMRV replicates efficiently in prostate epithelial cell lines specifically in LNCaP cells [3, 18]. In addition, the prostate cancer cell line 22Rv1 has been shown to be chronically infected with XMRV and produces highly infectious virus . Since host restriction factor A3G is able to restrict XMRV, the question is how XMRV replicates efficiently in these human prostate cell lines. There are at least three studies that have suggested that XMRV efficiently replicates in prostate epithelial cancer cell lines since these cells lack or express undetectable levels of A3G [13–16]. In this report, we demonstrate that prostate epithelial cell lines LNCaP and DU-145 express detectable levels of A3G by western blot analysis. We confirm the presence of A3G in LNCaP cells by mass spectrometry. We believe the results described in earlier reports on the absence of A3G in these cells may be due to the sensitivity of antibody used in their western blot analysis.
In the absence of Vif-like accessory proteins, retroviruses such as Human T cell lymphoma virus (HTLV) and Murine leukemia virus (MLV) have developed alternative mechanisms to evade host restriction by A3 proteins. A motif of HTLV nucleocapsid (NC) prevents packaging of A3G into the virion . Therefore exclusion of A3G has been proposed to be a common mechanism for Vif-deficient retroviruses to counteract A3G restriction. MLV virions have also been reported to exclude mA3 . XMRV has been demonstrated to package A3G , therefore a role for exclusion mechanism is unlikely. XMRV produced from LNCaP cells show signatures of hypermutation that are characteristics of A3F [15, 17]. Therefore, it is plausible XMRV is somehow resistant to A3G restriction in these cell types. XMRV may achieve this either by downregulating A3G levels or by evading A3G restriction. Given that MLV has been reported to inactivate mA3 by viral protease , a similar mechanism for XMRV cannot be ruled out. Furthermore, certain polymorphic alleles of A3G have been reported to increase the susceptibility to HIV infection . Therefore, we are investigating whether A3G produced from prostate epithelial cells have mutations that can be assigned for cell specific susceptibility to XMRV infection. The other mechanism that could possibly explain our results is that A3G remains as high-molecular-mass (HMM) ribonucleoprotein complex in prostate epithelial cells. It has been reported that A3G in resting CD4+ cells and monocytes are predominantly in its low-molecular-mass (LMM) active form making these cells refractory to HIV-1 infection [24, 25]. Conversion of LMM to the inactive HMM complex, when the CD4+ cells are activated or monocytes are differentiated into macrophages, makes these cells prone to HIV infection. Since HMM forms of A3G are reported to be enzymatically inactive, if A3G remains in HMM complex in prostate epithelial cells, it may not be able to restrict XMRV replication.
In summary, this report demonstrates the presence of A3G in prostate epithelial cell lines (LNCaP and DU145) that support efficient XMRV replication. Since XMRV packages A3G in its virions and lacks Vif-like accessory proteins, our findings on XMRV-induced downregulation of A3G may represent a new pathway by which retroviruses counteract antiviral effects of A3 proteins in human cells. Our data warrants further studies to decipher the mechanism by which XMRV may counteract restriction by A3 proteins.
We thank Dr. Sandra Ruscetti (NCI-Frederick) for the antibodies. We thank Dr. James Hildreth (UC-Davis) for technical guidance. This work is supported in part by grants to CD (R00DA024558, R03DA30896) from NIH, (UL1 RR024975) from the Vanderbilt CTSA Grant and (U54RR026140) Meharry Translational Research Center (MeTRC).
- Urisman A, Molinaro RJ, Fischer N, Plummer SJ, Casey G, Klein EA, Malathi K, Tubbs RR, Ganem D, Silverman RH, DeRisi JL: Identification of a novel gammaretrovirus in prostate tumors of patients homozygous for R462Q RNASEL variant. PLoSPathog 2006, 2: e25.Google Scholar
- Arnold RS, Makarova NV, Osunkoya AO, Suppiah S, Scott TA, Johnson NA, Bhosle SM, Liotta D, Hunter E, Marshall FF, et al.: XMRV infection in patients with prostatecancer: novel serologic assay and correlation with PCR andFISH. Urology 2010, 75: 755-761. 10.1016/j.urology.2010.01.038View ArticlePubMedGoogle Scholar
- Dong B, Kim S, Hong S, Das Gupta J, Malathi K, Klein EA, Ganem D, DeRisi JL, Chow SA, Silverman RH: An infectious retrovirus susceptible to an IFN antiviral pathway from human prostate tumors. ProcNatlAcadSci USA 2007, 104: 1655-60. 10.1073/pnas.0610291104View ArticleGoogle Scholar
- Schlaberg R, Choe DJ, Brown KR, Thaker HM, Singh IR: XMRV ispresent in malignant prostatic epithelium and is associated with prostate cancer, especially high-grade tumors. ProcNatlAcadSci USA 106: 16351-16356.Google Scholar
- Sabunciyan S, Mandelberg N, Rabkin CS, Yolken R, Viscidi R: No difference in antibody titers against xenotropicMLV related virus in prostate cancer cases and cancer-freecontrols. Mol Cell Probes 2010, 25: 134-136.View ArticleGoogle Scholar
- Fischer N, Hellwinkel O, Schulz C, Chun FK, Huland H, Aepfelbacher M, Schlomm T: Prevalence of human gammaretrovirus XMRV in sporadic prostate cancer. J ClinVirol 2008, 43: 277-83.Google Scholar
- Hohn O, Krause H, Barbarotto P, Niederstadt L, Beimforde N, Miller K, Kurth R, Bannert N: Lack of evidence for xenotropic murine leukemia virus-related virus (XMRV) in German prostate cancer patients. Retrovirology 2009, 6: 92. 10.1186/1742-4690-6-92PubMed CentralView ArticlePubMedGoogle Scholar
- Verhaegh GW, de Jong AS, Smit FP, et al.: Prevalence of human xenotropic murine leukemia virus-related gammaretrovirus (XMRV) in Dutch prostate cancer patients. Prostate 2010, 71: 415-20.View ArticlePubMedGoogle Scholar
- Aloia AL, Sfanos KS, Isaacs WB, et al.: XMRV: a new virus in prostate cancer? Cancer Res 2010, 70: 10028-33. 10.1158/0008-5472.CAN-10-2837PubMed CentralView ArticlePubMedGoogle Scholar
- Paprotka T, Delviks-Frankenberry KA, Cingöz O, Martinez A, Kung H-J, Tepper CG, Hu W-S, Fivash MJ Jr, Coffin MJ, Pathak VK: Recombinant Origin of the Retrovirus XMRV. Science 2011, in press. 31 May 2011/Page 1/10.1126/science.1205292Google Scholar
- Sheehy AM, Gaddis NC, Choi JD, Malim MH: Isolation of a human gene that inhibits HIV-1 infection and is suppressed by the viral Vif protein. Nature 2002, 418: 646-650. 10.1038/nature00939View ArticlePubMedGoogle Scholar
- Gabuzda DH, et al.: Role of vif in replication of human immunodeficiencyvirus type 1 in CD4 T lymphocytes. J Virol 1992, 66: 6489-6495.PubMed CentralPubMedGoogle Scholar
- Bogerd HP, Zhang FDBP, Cullen BR: Human APOBEC3proteins can inhibit xenotropic murine leukemia virus-related virus infectivity. Virology 2011, 410: 234-239. 10.1016/j.virol.2010.11.011PubMed CentralView ArticlePubMedGoogle Scholar
- Groom HC, Yap MW, Galao RP, Neil SJ, Bishop KN: Susceptibility of xenotropic murine leukemia virus-related virus (XMRV) to retroviral restriction factors. Proc Natl Acad Sci USA 2010, 107: 5166-5171. 10.1073/pnas.0913650107PubMed CentralView ArticlePubMedGoogle Scholar
- Paprotka T, et al.: Inhibition of xenotropic murine leukemia virusrelated virus by APOBEC3 proteins and antiviral drugs. J Virol 2010, 84: 5719-5729. 10.1128/JVI.00134-10PubMed CentralView ArticlePubMedGoogle Scholar
- Stieler K, Fischer N: Apobec 3 G efficiently reduces infectivity of the human exogenousgammaretrovirus XMRV. PLoS One 2010, 5: e11738. 10.1371/journal.pone.0011738PubMed CentralView ArticlePubMedGoogle Scholar
- Paprotka T, et al.: Inhibition of xenotropic murine leukemia virusrelatedvirus by APOBEC3 proteins in PBMCS. J Virol 2011, 85: 4888-4897. 10.1128/JVI.00046-11PubMed CentralView ArticlePubMedGoogle Scholar
- Rodriguez JJ, Goff SP: Xenotropic murine leukemia virus-related virus establishes an efficient spreading infection and exhibits enhanced transcriptional activity in prostate carcinoma cells. J Virol 2010,84(5):2556-62. 10.1128/JVI.01969-09PubMed CentralView ArticlePubMedGoogle Scholar
- Knouf EC, et al.: Multiple integrated copies and high-level production of the human retrovirus XMRV from 22Rv1 prostate carcinoma cells. J Virol 2009, 83: 7353-7356. 10.1128/JVI.00546-09PubMed CentralView ArticlePubMedGoogle Scholar
- Derse D, Hill SA, Princler G, Lloyd P, Heidecker G: Resistance of human T cell leukemia virus type 1 to APOBEC3G restriction is mediated by elements in nucleocapsid. Proc Natl Acad Sci USA 2007, 104: 2915-2920. 10.1073/pnas.0609444104PubMed CentralView ArticlePubMedGoogle Scholar
- Doehle BP, Schafer A, Wiegand HL, Bogerd HP, Cullen BR: Differential sensitivity of murine leukemia virus to APOBEC3-mediated inhibition is governed by virion exclusion. J Virol 2005, 79: 8201-8207. 10.1128/JVI.79.13.8201-8207.2005PubMed CentralView ArticlePubMedGoogle Scholar
- Abudu A, Takaori-Kondo A, Izumi T, Shirakawa K, Kobayashi M, et al.: Murine retrovirus escapes from murine APOBEC3 via two distinct novel mechanisms. Curr Biol 2006, 16: 1565-1570. 10.1016/j.cub.2006.06.055View ArticlePubMedGoogle Scholar
- An P, Bleiber G, Duggal P, Nelson G, May M, Mangeat B, Alobwede I, Trono D, Vlahov D, Donfield S, Goedert JJ, Phair J, Buchbinder S, O'Brien SJ, Telenti A, Winkler CA: APOBEC3G Genetic Variants and Their Influence on the Progression to AIDS. J Virol 2004, 78: 11070-11076. 10.1128/JVI.78.20.11070-11076.2004PubMed CentralView ArticlePubMedGoogle Scholar
- Chiu Y, Soros VB, Kreisberg J, Stopak K, Yonemoto Wes, Greene WC: Cellular APOBEC3G restricts HIV-1 infection in resting CD4 + T cells. Nature 2005, 435: 108-114. 10.1038/nature03493View ArticlePubMedGoogle Scholar
- Triques K, Stevenson M: Characterization of restrictions to human immunodeficiency virus type 1 infection of monocytes. J Virol 2004, 78: 5523-5527. 10.1128/JVI.78.10.5523-5527.2004PubMed CentralView ArticlePubMedGoogle Scholar
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