Encapsidation of APOBEC3G into HIV-1 virions involves lipid raft association and does not correlate with APOBEC3G oligomerization
© Khan et al; licensee BioMed Central Ltd. 2009
Received: 15 June 2009
Accepted: 03 November 2009
Published: 03 November 2009
The cellular cytidine deaminase APOBEC3G (A3G), when incorporated into the human immunodeficiency virus type 1 (HIV-1), renders viral particles non-infectious. We previously observed that mutation of a single cysteine residue of A3G (C100S) inhibited A3G packaging. In addition, several recent studies showed that mutation of tryptophan 127 (W127) and tyrosine 124 (Y124) inhibited A3G encapsidation suggesting that the N-terminal CDA constitutes a viral packaging signal in A3G. It was also reported that W127 and Y124 affect A3G oligomerization.
Here we studied the mechanistic basis of the packaging defect of A3G W127A and Y124A mutants. Interestingly, cell fractionation studies revealed a strong correlation between encapsidation, lipid raft association, and genomic RNA binding of A3G. Surprisingly, the presence of a C-terminal epitope tag affected lipid raft association and encapsidation of the A3G W127A mutant but had no effect on wt A3G encapsidation, lipid raft association, and interaction with viral genomic RNA. Mutation of Y124 abolished A3G encapsidation irrespective of the presence or absence of an epitope tag. Contrasting a recent report, our co-immunoprecipitation studies failed to reveal a correlation between A3G oligomerization and A3G encapsidation. In fact, our W127A and Y124A mutants both retained the ability to oligomerize.
Our results confirm that W127 and Y124 residues in A3G are important for encapsidation into HIV-1 virions and our data establish a novel correlation between genomic RNA binding, lipid raft association, and viral packaging of A3G. In contrast, we were unable to confirm a role of W127 and Y124 in A3G oligomerization and we thus failed to confirm a correlation between A3G oligomerization and virus encapsidation.
APOBEC3G (A3G) is a cellular cytidine deaminase with potent antiretroviral activity that severely limits replication of vif-defective HIV-1 in human cells . A3G is expressed in most if not all natural human HIV-1 target cells; yet HIV-1 efficiently infects humans and has caused a worldwide pandemic. This ability of HIV-1 to infect and replicate in A3G-positive human cells is made possible by the viral accessory protein Vif, which was found to prevent the packaging of A3G into progeny virions. Inhibition of A3G packaging is accomplished either by proteasome-mediated degradation of A3G or by other degradation-independent mechanisms (reviewed in ). Inhibition of A3G encapsidation may also require Vif dimerization since peptide antagonists to Vif dimerization blocked A3G packaging without affecting its intracellular stability .
The antiviral effect of A3G generally requires encapsidation of the deaminase into viral particles. Interestingly, the antiviral effects of A3G are not limited to HIV-1 but extend to other retroviruses including murine leukemia virus, mouse mammary tumor virus, simian immunodeficiency virus, equine infectious anemia virus, and hepatitis B virus (for review see ). Packaging of A3G into such diverse viruses suggests that virus encapsidation is either relatively nonspecific or involves signals shared by these viruses. Interestingly, although A3G selectively targets single stranded DNA for deamination it also binds RNA. RNA binding of A3G has been shown to contribute to virus encapsidation [4–11]. A3G also interacts with the NC component of the viral Gag precursor protein [7, 12–21]. This interaction likely also contributes to the packaging of A3G into viral particles. In vitro studies using purified recombinant NC and A3G found that the two proteins do not competitively bind RNA but instead form an RNA-protein ternary complex .
Several reports have investigated domains in A3G required for packaging into HIV-1 virions. We and others have recently reported that mutations in the A3G catalytic domain 1 (CD1) can impair A3G packaging [21, 22]. Characterization of in-frame deletion mutants implicated a linker region located C-terminal to the CD1 domain (residues 121-161) as critical for A3G packaging into HIV-1 virus-like particles [12, 20]. These findings were supported by other studies that identified residues 122 to 127 in the linker domain as important for A3G encapsidation [9, 23–26]) It is interesting to note that the adjacent D128 plays an important role in the species specific sensitivity of A3G to Vif [27–30]. Thus, the N-terminal linker region appears to be an important contact point for Vif as well as a requirement for A3G encapsidation. However, there is no conclusive evidence that these regions in A3G constitute direct Vif and/or Gag binding sites as of yet. It is equally possible that these regions impose conformational constraints on the protein that indirectly affect A3G encapsidation or modulate binding of Vif to other regions of the protein. In support of the latter possibility, Stenglein et al. have recently found that the W127A mutation has profound effects on A3G's intracellular localization only in conjunction with simultaneous mutation of Y19 . Based on structural predictions, W127 is located at the protein surface [26, 31]. and might therefore be available for a variety of functions including protein-protein and protein-nucleic acid interactions. Indeed, the packaging defect of the A3G W127A mutant was explained by an inability of this mutant to interact with 7SL RNA [9, 24]. More recently, the packaging defect of W127A and Y124A mutants was correlated with a defect in A3G oligomerization and the authors proposed that RNA-dependent oligomerization of APOBEC3G was required for restricting HIV-1 .
Here we further characterized the role of W127 and Y124 for the packaging of A3G into HIV-1 virions and for A3G oligomerization. Consistent with previous reports we found that packaging of A3G-HA was severely affected by the W127A mutation. Similarly, packaging of Myc epitope-tagged A3G-Myc W127A was severely restricted suggesting that the packaging defect imposed by the W127A mutation is not epitope tag specific. Of note, the effect of the W127A mutation on virus encapsidation was much less severe in the context of untagged A3G. This is surprising and implies that the effects of mutations around position W127 are sensitive to and exacerbated by changes at the C-terminus of the protein. In contrast, mutation of Y124A imposed a severe packaging defect irrespective of the presence or absence of an epitope tag. A3G-HA was previously found to associate with cellular raft structures . Interestingly, our results identified a novel correlation between A3G raft association and virus encapsidation. We analyzed a total of nine A3G variants and found that all packaging competent A3G variants associated with lipid rafts while all packaging incompetent A3G variants failed to do so. We further found that all packaging competent A3G variants interacted with genomic viral RNA as well as 7SL RNA while all packaging incompetent variants interacted with 7SL RNA but failed to interact with viral genomic RNA. Finally, all of our A3G variants analyzed in this study retained the ability to oligomerize irrespective of whether the A3G variant was packaging competent or not. Thus, our data clearly establish a positive correlation between packaging competence of A3G and the ability to associate with lipid rafts and to interact with viral genomic RNA. In contrast, our data failed to verify a correlation between A3G oligomerization and packaging competence. Finally, our results suggest that the presence of C-terminal epitope tags in A3G can impose conformational constraints on A3G that appear to be functionally inconsequential in the context of wild type protein but can exacerbate defects induced by changes to other regions of the protein such as mutation of W127.
The vif-defective molecular clone pNL4-3Vif(-) was used for the production of virus. Wild type human A3G carrying a C-terminal Myc epitope tag was described previously . For the expression of untagged human A3G, a stop codon was introduced into pcDNA-A3G-Myc by PCR-directed mutagenesis as reported elsewhere . Mutation of tryptophan residue W127 and tyrosine residue Y124 to alanine in Myc-tagged and untagged human A3G was accomplished by PCR-based mutagenesis of pcDNA-APO3G-Myc and pcDNA-APO3G vector, respectively. The presence of the desired mutation was verified by sequence analysis. Both tagged and untagged A3G were detected by the A3G-specific ApoC17 rabbit polyclonal antibody and were distinguishable by their different mobilities in the gel. Plasmids pA3G, pA3G-HA, pA3G W127A and pA3G-HA W127A expressing untagged and C-terminally HA-tagged A3G wt and W127A mutants in the backbone of pCMV4-HA were a gift of Michael Malim .
Tissue culture and transfection
HeLa cells were propagated in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (FBS). For transfection, HeLa cells were grown in 25 cm2 flasks to about 80% confluence. Cells were transfected using LipofectAMINE PLUS (Invitrogen Crop., Carlsbad CA) following the manufacturer's recommendations. A total of 5 to 6 μg of plasmid DNA per 25 cm2 flasks (~5 × 106 cells) was used. Total amounts of transfected DNA were kept constant in all samples of any given experiment by adding empty vector DNA (pUC18 or pcDNA3.1(-)MycHis) as appropriate. Unless stated otherwise, cells were harvested 24 h post-transfection.
A3G was identified using a polyclonal rabbit serum against a synthetic peptide comprising the 17 C-terminal residues of A3G (anti-ApoC17; available through the NIH AIDS Research and Reagent Program, Cat # 10082). Serum from an HIV-positive patient (APS) was used to detect HIV-1-specific capsid (CA) proteins. Tubulin was identified using a monoclonal antibody to α-tubulin (Sigma-Aldrich, Inc., St. Louis MO; Cat # T9026). For immunoprecipitation of tagged and untagged A3G, polyclonal ApoC17 antibody was used. Raft associated marker protein caveolin was identified by polyclonal anti-caveolin antibody (BD Bioscience Pharmingen, San Diego CA; Cat # 610060). Transferrin receptor (TfR) was included as a non raft marker protein and was identified using a TfR-specific monoclonal antibody (BD Bioscience Pharmingen, San Diego CA; Cat # 612125).
Preparation of virus stocks
Virus stocks were prepared by transfection of HeLa cells with appropriate plasmid DNAs of pNL4-3Vif(-) in the presence of tagged and untagged variants of wild type and mutant (W127A, Y124A) A3G as indicated in the text. Virus-containing supernatants were harvested 24 h after transfection. Cellular debris was removed by centrifugation (3 min, 3,000 × g) and clarified supernatants were filtered (0.45 μm) to remove residual cellular contaminants. For determination of viral infectivity, unconcentrated filtered supernatants were used for the infection of LuSIV indicator cells. For immunoblot analysis of viral protein, virus-containing supernatants (7 ml) were concentrated by ultracentrifugation through 4 ml of 20% sucrose in phosphate-buffered saline (PBS) as described previously .
To determine viral infectivity, virus stocks were normalized for equal levels of reverse transcriptase activity and used to infect LuSIV cells (5 × 105) in a 24-well plate in a total volume of 1.2 to 1.4 ml. LuSIV cells are derived from CEMx174 cells and contain a luciferase indicator gene under the control of the SIVmac239 long terminal repeat . These cells were obtained from Janice Clements through the NIH AIDS Research and Reference Reagent Program (catalog # 5460) and were maintained in complete RPMI 1640 medium supplemented with 10% FBS and hygromycin B (300 μg/ml). Cells were infected for 24 h at 37°C. Cells were then harvested and lysed in 150 μl of Promega 1× reporter lysis buffer (Promega Crop., Madison WI). To determine the luciferase activity in the lysates, 50 μl of each lysate was combined with luciferase substrate (Promega. Corp., Madison WI) by automatic injection and light emission was measured for 10 seconds at room temperature in a luminometer (Opticomp II; MGM instruments, Hamden CT).
For immunoblot analysis of intracellular proteins, whole-cell lysates were prepared as follows. Cells were washed once with PBS, suspended in PBS (400 μl/107 cells), and mixed with an equal volume of sample buffer (4% sodium dodecyl sulfate [SDS], 125 mM Tris-HCL, pH 6.8, 10% 2-mercaptoethanol, 10% glycerol and 0.002% bromophenol blue). Proteins were solubilized by boiling for 10 to 15 min at 95°C, with occasional vortexing of the samples to shear cellular DNA. Residual insoluble material was removed by centrifugation (2 min, 15,000 rpm, in an Eppendorf Minifuge). For immunoblot analysis of virus-associated proteins, concentrated viral pellets were suspended in a 1:1 mixture of PBS and sample buffer and boiled. Cell lysates and viral extracts were subjected to SDS-polyacrylamide gel electrophoresis; proteins were transferred to polyvinylidene diflouride membranes and reacted with appropriate antibodies as described in the text. Membranes were then incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies (GE Healthcare Biosciences, Piscataway NJ) and visualized by enhanced chemiluminescence (GE Healthcare Biosciences).
For immunoprecipitation of tagged and untagged A3G, A3G W127A, and A3G Y124A, lysates of transfected cells were prepared as follows. Cells were washed once with PBS and lysed in 300 μl of lysis buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 0.5% Triton X-100). Cell extracts were clarified at 13,000 × g for 3 min, and the supernatant was incubated on a rotating wheel for 1 h at 4°C with protein A-Sepharose coupled with anti-ApoC17 antibody. Immune complexes were washed three times with 50 mM Tris, 300 mM NaCl, and 0.1% Triton X-100, pH 7.4. Bound proteins were eluted from beads by heating in sample buffer for 5 min at 96°C and analyzed by immunoblotting.
HeLa cells were transfected with 2.5 μg each of vectors expressing untagged or C-terminally Myc tagged A3G proteins in various combinations. Cells were lysed in 600 ml lysis buffer (0.5% Triton X-100, 287 mM NaCl, 2.68 mM KCl, 1.47 mM KH2PO4, Na2HPO4, pH 7.2) as described . Lysates were immunoprecipitated with a Myc-specific monoclonal antibody (clone 9E10; Sigma-Aldrich, Inc., St. Louis MO; Cat # M 4439) as described above. Immunoprecipitates were subjected to immunoblot analysis using an A3G-specific rabbit polyclonal antibody (Apo-C17).
Membrane floatation analysis (raft association)
Raft association of A3G was assessed by membrane floatation analyses essentially as described by Ono et al . HeLa cells were transfected with 5 μg of wild type and mutant A3G expression constructs DNA (pcDNA-APO3G, pcDNA-APO3G-Myc, pcDNA-APO3G-W127A, pcDNA-APO3G-W127A-Myc, and pcDNA-APO3G-C100S-Myc, respectively). Cells were harvested 20 h later by scraping and washed three times with ice-cold PBS. Cells were pelleted (2,000 × g for 2 min) and resuspended in 300 μl of 10 mM Tris-HCl pH 7.5 supplemented with 4 mM EDTA and Complete™ protease inhibitor cocktail (Roche Diagnostics Corp., Indianapolis IN). After 10 min incubation on ice cells were sonicated for 10 sec and centrifuged for 3 min at 2,000 × g at 4°C in a microcentrifuge to remove insoluble material and nuclei. The postnuclear supernatants (120 μl) were mixed with 120 μl of TNE lysis buffer (100 mM Tris-HCl, 600 mM NaCl and 16 mM EDTA) containing 0.5% Triton X-100 and incubated on ice for 20 min. A total of 200 μl of each lysate was mixed with 1 ml of 85.5% sucrose (w/v) in TNE lysis buffer, placed at the bottom of ultracentrifuge tubes, and overlaid with 2.5 ml of 65% (w/v) sucrose and 1.5 ml of 10% sucrose (w/v) in TNE lysis buffer. The samples were centrifuged at 4°C in a SW55 rotor for 16 hours at 35,000 rpm to obtain Triton X-100 resistant and sensitive fractions. Ten equal fractions (500 μl each) were collected from the top, mixed with 4× sample buffer (180 μl) and boiled. Samples were analyzed by immunoblotting.
Total cellular RNA was extracted from untransfected and transfected HeLa cells using the RNeasy RNA extraction kit (QIAGEN, Valencia CA) following the manufacturer's instructions. To isolate RNA form immune complexes, beads were washed three times with RNA-protein binding buffer (20 mM HEPES, 25 mM KCl, 7 mM 2-Mercaptoehanol, 5% Glycerol and 0.1% NP-40). RNA was then extracted using the RNeasy RNA extraction kit. For isolation of genomic RNA precipitated with the A3G complex, vif-defective HIV-1 proviral vector DNA (1 μg) was co-transfected into HeLa cells with A3G vectors (4 μg) as indicated in the text. RNA was then extracted from the immunocomplexes as above.
qRT-PCR was performed using the one tube SYBR green method as per manufacturer instruction (AB Biosystems, Warrington UK). Briefly, each 16 μl reaction mixture contained 0.08 μl of Reverse Transcriptase, 0.04 μl of RNase inhibitor, 300 μM of forward and 50 μM of reverse specific primers, 8 μl of 2× SYBR green PCR Master Mix, 2.6 μl of RNase-free water, and 5 μl of template RNA. RNaseA treated RNA from A3G wt samples were used as a negative control. The reactions were performed on a AB Biosystems 7300 Real Time PCR System (AB Biosystems) using the following conditions: 48°C for 30 min followed by 95°C for 10 min and then 40 cycles of 95°C for 15 s and 60°C for 1 min with a dissociation protocol. The target sequences amplified by the SYBR green method used the following primer pairs: 7SL RNA, forward (5'-CCCGGGAGGTCACCATATT-3'), reverse (5'-CTGTAGTCCCAGCTACTCG-3'); HIV-1 genomic RNA, forward (5'TCAGCATTATCAGAAGGAGCCACC-3'), reverse (5'-TCATCCATCCTATTTGTTCCTGAAG-3').
Mutation of W127 induces a packaging defect in A3G that is exacerbated by C-terminal epitope tags
To address potential effects of epitope tags on packaging of A3G, we performed a side-by-side comparison of untagged and epitope tagged A3G wt, A3G W127A, as well as A3G Y124A. For the Y124A mutant the presence or absence of a C-terminal Myc tag was investigated; A3G W127A constructs encoding either a C-terminal Myc tag or an HA tag were analyzed. Amounts of transfected A3G vectors were adjusted as described in the legend to figure 1 to minimize differences in expression or stability of the proteins (Fig. 1A, cell). All DNAs were cotransfected into HeLa cells together with an equal amount of vif-defective proviral DNA (pNL4-3Vif(-)) and total amounts of transfected DNA were kept constant. Surprisingly, epitope-tagged A3G W127A variants were much less efficiently packaged into virions than their untagged counterparts (Fig. 1B, compare lanes 5 & 7 to lanes 6 & 8). In contrast, A3G Y124A was packaging incompetent irrespective of the present or absence of a C-terminal Myc tag (Fig. 1B, lanes 9 & 10). Thus, the presence of a C-terminal epitope tag, irrespective of its nature (i.e Myc versus HA), reduced the packaging efficiency of W127A mutants by 40- to 60-fold when compared to untagged A3G wt.
Packaging of A3G correlates with lipid raft association
A3G packaging competence correlates with the ability to interact with viral genomic RNA
Lack of correlation between A3G oligomerization and packaging competence
We previously reported that mutation of C97 in the N-terminal enzymatically inactive deaminase domain of A3G affected oligomerization of the protein but did not abolish packaging or antiviral activity . In contrast, a more recent study concluded that RNA-dependent oligomerization of A3G is required for packaging and for restriction of HIV-1 . This conclusion is based on the observation that the packaging defective A3G Y124 and W127 mutants failed to interact with A3G wt in co-immunoprecipitation studies.
The mechanism of A3G encapsidation into HIV-1 virions has attracted significant attention since it offers a potential target for therapeutic interference with virus replication. There is increasing evidence that encapsidation of A3G into virions requires interactions with the viral nucleocapsid domain in the Gag precursor and involves RNA although the nature of the RNA involved in A3G packaging, i.e. cellular versus viral, remains under investigation [8, 9, 12–15, 18–20, 38, 41–43]. There is only limited information concerning sequences in A3G that are necessary and sufficient for viral encapsidation. Several studies implicated the N-terminal catalytic domain (CD1) in A3G in RNA interaction and virus encapsidation [5, 21, 22]. Other studies found that sequences in the N-terminal linker domain downstream of the CD1 domain encompassing residues 122 to 127 were critical for A3G packaging [9, 12, 20, 23, 25, 26]. However, it remains unclear whether this latter domain represents a direct contact point for protein-protein or protein-RNA interactions or simply represents a conformationally sensitive area in the protein.
Our current study focused on two amino acid residue, W127 and Y124, in A3G to demonstrate that the requirements for A3G packaging are complex and can be influenced by the presence or absence of terminal epitope tags. Our finding that untagged A3G W127A was packaged 10 times more efficient than Myc- or HA-tagged A3G W127A was surprising and unexpected since the interfering epitope tags were not located adjacent to residue W127 but were more than 250 amino acids away at the C-terminus of the protein. There is currently no structure of full-length A3G available. However, computer modelling suggests that W127 is located at the surface of the protein . It is therefore possible that in 3-dimensional space the C-terminus of A3G is in close proximity to the N-terminal linker region surrounding residue W127. Therefore, changes in this region could, in the context of an epitope-tag, induce changes in the protein resulting in mislocalization in the cell - as evidenced by differential raft association - and culminating in the exclusion of A3G from virions. Importantly, the epitope tag effect was not tag specific since identical results were obtained with C-terminal Myc and HA tags. Based on these data we consider it possible that W127 in A3G is critical for proper protein folding/conformation and/or cellular localization of the protein rather than representing a motif required for viral encapsidation.
Our data reveal an interesting correlation between A3G's propensity to associate with lipid raft structures, viral RNA interaction, and packaging competence. The reason for the inability of the epitope tagged A3G W127A mutants as well as tagged and untagged Y124A mutants to associate with lipid rafts and with viral RNA remains to be investigated. Since A3G is not a membrane protein it is likely that its affinity to raft structures is due to an interaction with other raft associated host factors. Our floatation studies were done in the absence of viral proteins, ruling out the possible contribution of Gag protein in the raft targeting of A3G. Thus, the lack of raft association could be indicative of altered intracellular localization or trafficking of A3G or could be due to a conformational change resulting in the loss of protein-protein interaction.
We thank Amy Andrew for critical comments on the manuscript and Melissa Gilden and Sandrine Opi for plasmid construction and help with reagents. We are grateful to Michael Malim for providing pA3G, pA3G W127A, pA3G-HA, and pA3G-HA W127A vectors. This work was supported by a Grant from the NIH Intramural AIDS Targeted Antiviral Program to K.S. and by the Intramural Research Program of the NIH, NIAID.
- 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/nature00939.View ArticlePubMedGoogle Scholar
- Goila-Gaur R, Strebel K: HIV-1 Vif, APOBEC, and intrinsic immunity. Retrovirology. 2008, 5: 51-10.1186/1742-4690-5-51.PubMed CentralView ArticlePubMedGoogle Scholar
- Miller JH, Presnyak V, Smith HC: The dimerization domain of HIV-1 viral infectivity factor Vif is required to block virion incorporation of APOBEC3G. Retrovirology. 2007, 4: 81-10.1186/1742-4690-4-81.PubMed CentralView ArticlePubMedGoogle Scholar
- Jarmuz A, Chester A, Bayliss J, Gisbourne J, Dunham I, Scott J, Navaratnam N: An anthropoid-specific locus of orphan C to U RNA-editing enzymes on chromosome 22. Genomics. 2002, 79: 285-296. 10.1006/geno.2002.6718.View ArticlePubMedGoogle Scholar
- Iwatani Y, Takeuchi H, Strebel K, Levin JG: Biochemical Activities of Highly Purified, Catalytically Active Human APOBEC3G: Correlation with Antiviral Effect. J Virol. 2006, 80: 5992-6002. 10.1128/JVI.02680-05.PubMed CentralView ArticlePubMedGoogle Scholar
- Yu Q, Konig R, Pillai S, Chiles K, Kearney M, Palmer S, Richman D, Coffin JM, Landau NR: Single-strand specificity of APOBEC3G accounts for minus-strand deamination of the HIV genome. Nat Struct Mol Biol. 2004, 11: 435-442. 10.1038/nsmb758.View ArticlePubMedGoogle Scholar
- Khan MA, Kao S, Miyagi E, Takeuchi H, Goila-Gaur R, Opi S, Gipson CL, Parslow TG, Ly H, Strebel K: Viral RNA is required for the association of APOBEC3G with human immunodeficiency virus type 1 nucleoprotein complexes. J Virol. 2005, 79: 5870-5874. 10.1128/JVI.79.9.5870-5874.2005.PubMed CentralView ArticlePubMedGoogle Scholar
- Khan MA, Goila-Gaur R, Opi S, Miyagi E, Takeuchi H, Kao S, Strebel K: Analysis of the contribution of cellular and viral RNA to the packaging of APOBEC3G into HIV-1 virions. Retrovirology. 2007, 4: 48-10.1186/1742-4690-4-48.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang T, Tian C, Zhang W, Luo K, Sarkis PT, Yu L, Liu B, Yu Y, Yu XF: 7SL RNA mediates virion packaging of the antiviral cytidine deaminase APOBEC3G. J Virol. 2007, 81: 13112-13124. 10.1128/JVI.00892-07.PubMed CentralView ArticlePubMedGoogle Scholar
- Tian C, Wang T, Zhang W, Yu XF: Virion packaging determinants and reverse transcription of SRP RNA in HIV-1 particles. Nucleic Acids Res. 2007, 35: 7288-7302. 10.1093/nar/gkm816.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang T, Tian C, Zhang W, Sarkis PT, Yu XF: Interaction with 7SL RNA but not with HIV-1 genomic RNA or P bodies is required for APOBEC3F virion packaging. J Mol Biol. 2008, 375: 1098-1112. 10.1016/j.jmb.2007.11.017.View ArticlePubMedGoogle Scholar
- Cen S, Guo F, Niu M, Saadatmand J, Deflassieux J, Kleiman L: The interaction between HIV-1 Gag and APOBEC3G. J Biol Chem. 2004, 279: 33177-33184. 10.1074/jbc.M402062200.View ArticlePubMedGoogle Scholar
- Schafer A, Bogerd HP, Cullen BR: Specific packaging of APOBEC3G into HIV-1 virions is mediated by the nucleocapsid domain of the gag polyprotein precursor. Virology. 2004, 328: 163-168. 10.1016/j.virol.2004.08.006.View ArticlePubMedGoogle Scholar
- Alce TM, Popik W: APOBEC3G is incorporated into virus-like particles by a direct interaction with HIV-1 Gag nucleocapsid protein. J Biol Chem. 2004, 279: 34083-34086. 10.1074/jbc.C400235200.View ArticlePubMedGoogle Scholar
- Zennou V, Perez-Caballero D, Gottlinger H, Bieniasz PD: APOBEC3G incorporation into human immunodeficiency virus type 1 particles. J Virol. 2004, 78: 12058-12061. 10.1128/JVI.78.21.12058-12061.2004.PubMed CentralView ArticlePubMedGoogle Scholar
- Mariani R, Chen D, Schrofelbauer B, Navarro F, Konig R, Bollman B, Munk C, Nymark-McMahon H, Landau NR: Species-specific exclusion of APOBEC3G from HIV-1 virions by Vif. Cell. 2003, 114: 21-31. 10.1016/S0092-8674(03)00515-4.View ArticlePubMedGoogle Scholar
- Mangeat B, Turelli P, Caron G, Friedli M, Perrin L, Trono D: Broad antiretroviral defence by human APOBEC3G through lethal editing of nascent reverse transcripts. Nature. 2003, 424: 99-103. 10.1038/nature01709.View ArticlePubMedGoogle Scholar
- Burnett A, Spearman P: APOBEC3G Multimers Are Recruited to the Plasma Membrane for Packaging into Human Immunodeficiency Virus Type 1 Virus-Like Particles in an RNA-Dependent Process Requiring the NC Basic Linker. J Virol. 2007, 81: 5000-5013. 10.1128/JVI.02237-06.PubMed CentralView ArticlePubMedGoogle Scholar
- Douaisi M, Dussart S, Courcoul M, Bessou G, Vigne R, Decroly E: HIV-1 and MLV Gag proteins are sufficient to recruit APOBEC3G into virus-like particles. Biochem Biophys Res Commun. 2004, 321: 566-573. 10.1016/j.bbrc.2004.07.005.View ArticlePubMedGoogle Scholar
- Luo K, Liu B, Xiao Z, Yu Y, Yu X, Gorelick R, Yu XF: Amino-terminal region of the human immunodeficiency virus type 1 nucleocapsid is required for human APOBEC3G packaging. J Virol. 2004, 78: 11841-11852. 10.1128/JVI.78.21.11841-11852.2004.PubMed CentralView ArticlePubMedGoogle Scholar
- Navarro F, Bollman B, Chen H, Konig R, Yu Q, Chiles K, Landau NR: Complementary function of the two catalytic domains of APOBEC3G. Virology. 2005, 333: 374-386. 10.1016/j.virol.2005.01.011.View ArticlePubMedGoogle Scholar
- Opi S, Takeuchi H, Kao S, Khan MA, Miyagi E, Goila-Gaur R, Iwatani Y, Levin JG, Strebel K: Monomeric APOBEC3G is catalytically active and has antiviral activity. J Virol. 2006, 80: 4673-4682. 10.1128/JVI.80.10.4673-4682.2006.PubMed CentralView ArticlePubMedGoogle Scholar
- Huthoff H, Malim MH: Identification of Amino Acid Residues in APOBEC3G Required for Regulation by Human Immunodeficiency Virus Type 1 Vif and Virion Encapsidation. J Virol. 2007, 81: 3807-3815. 10.1128/JVI.02795-06.PubMed CentralView ArticlePubMedGoogle Scholar
- Bach D, Peddi S, Mangeat B, Lakkaraju A, Strub K, Trono D: Characterization of APOBEC3G binding to 7SL RNA. Retrovirology. 2008, 5: 54-10.1186/1742-4690-5-54.PubMed CentralView ArticlePubMedGoogle Scholar
- Gooch BD, Cullen BR: Functional domain organization of human APOBEC3G. Virology. 2008, 379: 118-124. 10.1016/j.virol.2008.06.013.PubMed CentralView ArticlePubMedGoogle Scholar
- Zhang KL, Mangeat B, Ortiz M, Zoete V, Trono D, Telenti A, Michielin O: Model Structure of Human APOBEC3G. PLoS ONE. 2007, 2: e378-10.1371/journal.pone.0000378.PubMed CentralView ArticlePubMedGoogle Scholar
- Bogerd HP, Doehle BP, Wiegand HL, Cullen BR: A single amino acid difference in the host APOBEC3G protein controls the primate species specificity of HIV type 1 virion infectivity factor. Proc Natl Acad Sci USA. 2004, 101: 3770-3774. 10.1073/pnas.0307713101.PubMed CentralView ArticlePubMedGoogle Scholar
- Schrofelbauer B, Chen D, Landau NR: A single amino acid of APOBEC3G controls its species-specific interaction with virion infectivity factor (Vif). Proc Natl Acad Sci USA. 2004, 101: 3927-3932. 10.1073/pnas.0307132101.PubMed CentralView ArticlePubMedGoogle Scholar
- Mangeat B, Turelli P, Liao S, Trono D: A single amino acid determinant governs the species-specific sensitivity of APOBEC3G to Vif action. J Biol Chem. 2004, 279: 14481-14483. 10.1074/jbc.C400060200.View ArticlePubMedGoogle Scholar
- Xu H, Svarovskaia ES, Barr R, Zhang Y, Khan MA, Strebel K, Pathak VK: A single amino acid substitution in human APOBEC3G antiretroviral enzyme confers resistance to HIV-1 virion infectivity factor-induced depletion. Proc Natl Acad Sci USA. 2004, 101: 5652-5657. 10.1073/pnas.0400830101.PubMed CentralView ArticlePubMedGoogle Scholar
- Stenglein MD, Matsuo H, Harris RS: Two regions within the amino-terminal half of APOBEC3G cooperate to determine cytoplasmic localization. J Virol. 2008, 82: 9591-9599. 10.1128/JVI.02471-07.PubMed CentralView ArticlePubMedGoogle Scholar
- Huthoff H, Autore F, Gallois-Montbrun S, Fraternali F, Malim MH: RNA-dependent oligomerization of APOBEC3G is required for restriction of HIV-1. PLoS Pathog. 2009, 5: e1000330-10.1371/journal.ppat.1000330.PubMed CentralView ArticlePubMedGoogle Scholar
- Karczewski MK, Strebel K: Cytoskeleton association and virion incorporation of the human immunodeficiency virus type 1 Vif protein. J Virol. 1996, 70: 494-507.PubMed CentralPubMedGoogle Scholar
- Kao S, Khan MA, Miyagi E, Plishka R, Buckler-White A, Strebel K: The human immunodeficiency virus type 1 Vif protein reduces intracellular expression and inhibits packaging of APOBEC3G (CEM15), a cellular inhibitor of virus infectivity. J Virol. 2003, 77: 11398-11407. 10.1128/JVI.77.21.11398-11407.2003.PubMed CentralView ArticlePubMedGoogle Scholar
- Opi S, Kao S, Goila-Gaur R, Khan MA, Miyagi E, Takeuchi H, Strebel K: Human immunodeficiency virus type 1 Vif inhibits packaging and antiviral activity of a degradation-resistant APOBEC3G variant. J Virol. 2007, 81: 8236-8246. 10.1128/JVI.02694-06.PubMed CentralView ArticlePubMedGoogle Scholar
- Roos JW, Maughan MF, Liao Z, Hildreth JE, Clements JE: LuSIV cells: a reporter cell line for the detection and quantitation of a single cycle of HIV and SIV replication. Virology. 2000, 273: 307-315. 10.1006/viro.2000.0431.View ArticlePubMedGoogle Scholar
- Ono A, Freed EO: Plasma membrane rafts play a critical role in HIV-1 assembly and release. Proc Natl Acad Sci USA. 2001, 98: 13925-13930. 10.1073/pnas.241320298.PubMed CentralView ArticlePubMedGoogle Scholar
- Bogerd HP, Cullen BR: Single-stranded RNA facilitates nucleocapsid: APOBEC3G complex formation. RNA. 2008, 14: 1228-1236. 10.1261/rna.964708.PubMed CentralView ArticlePubMedGoogle Scholar
- Wang T, Zhang W, Tian C, Liu B, Yu Y, Ding L, Spearman P, Yu XF: Distinct viral determinants for the packaging of human cytidine deaminases APOBEC3G and APOBEC3C. Virology. 2008, 377: 71-79. 10.1016/j.virol.2008.04.012.PubMed CentralView ArticlePubMedGoogle Scholar
- Zhang L, Li X, Ma J, Yu L, Jiang J, Cen S: The incorporation of APOBEC3 proteins into murine leukemia viruses. Virology. 2008, 378: 69-78. 10.1016/j.virol.2008.05.006.View ArticlePubMedGoogle Scholar
- Goila-Gaur R, Khan MA, Miyagi E, Kao S, Strebel K: Targeting APOBEC3A to the viral nucleoprotein complex confers antiviral activity. Retrovirology. 2007, 4: 61-10.1186/1742-4690-4-61.PubMed CentralView ArticlePubMedGoogle Scholar
- Svarovskaia ES, Xu H, Mbisa JL, Barr R, Gorelick RJ, Ono A, Freed EO, Hu WS, Pathak VK: Human apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) is incorporated into HIV-1 virions through interactions with viral and nonviral RNAs. J Biol Chem. 2004, 279: 35822-35828. 10.1074/jbc.M405761200.View ArticlePubMedGoogle Scholar
- Soros VB, Yonemoto W, Greene WC: Newly synthesized APOBEC3G is incorporated into HIV virions, inhibited by HIV RNA, and subsequently activated by RNase H. PLoS Pathog. 2007, 3: e15-10.1371/journal.ppat.0030015.PubMed CentralView ArticlePubMedGoogle Scholar
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