Differential Contributions of Ubiquitin-Modified APOBEC3G Lysine Residues to HIV-1 Vif-Induced Degradation.
Apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (A3G) is a host restriction factor that impedes HIV-1 replication. Viral integrity is salvaged by HIV-1 virion infectivity factor (Vif), which mediates A3G polyubiquitination and subsequent cellular depletion. Previous studies have implied that A3G polyubiquitination is essential for Vif-induced degradation. However, the contribution of polyubiquitination to the rate of A3G degradation remains unclear. Here, we show that A3G polyubiquitination is essential for degradation. Inhibition of ubiquitin-activating enzyme E1 by PYR-41 or blocking the formation of ubiquitin chains by over-expressing the lysine to arginine mutation of ubiquitin K48 (K48R) inhibited A3G degradation. Our A3G mutagenesis study showed that lysine residues 297, 301, 303, and 334 were not sufficient to render lysine-free A3G sensitive to Vif-mediated degradation. Our data also confirm that Vif could induce ubiquitin chain formation on lysine residues interspersed throughout A3G. Notably, A3G degradation relied on the lysine residues involved in polyubiquitination. Although A3G and the A3G C-terminal mutant interacted with Vif and were modified by ubiquitin chains, the latter remained more resistant to Vif-induced degradation. Furthermore, the A3G C-terminal mutant, but not the N-terminal mutant, maintained potent antiviral activity in the presence of Vif. Taken together, our results suggest that the location of A3G ubiquitin modification is a determinant for Vif-mediated degradation, implying that in addition to polyubiquitination, other factors may play a key role in the rate of A3G degradation.
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA.
Tiffany
T
Turner
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA.
Qiujia
Q
Shao
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA; National Engineering Laboratory for AIDS Vaccine, School of Life Science, Jilin University, Changchun, Jilin Province 130000, People's Republic of China.
Weiran
W
Wang
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA.
Yudi
Y
Wang
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA.
Chenliang
C
Wang
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA.
Ballington
B
Kinlock
Center for AIDS Health Disparities Research, Department of Microbiology and Immunology, Meharry Medical College, Nashville, TN 37208, USA. Electronic address: bliu@mmc.edu.
Bindong
B
Liu
D000071480:APOBEC-3G Deaminase / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D025801:Ubiquitin / Q000378:metabolism*; D054320:vif Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
D025801:Ubiquitin; D054320:vif Gene Products, Human Immunodeficiency Virus; C514847:vif protein, Human immunodeficiency virus 1; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D008239:Lysine
APOBEC3G lysine free mutant; host restriction factor; polyubiquitination; proteasomal degradation; ubiquitin-activating enzyme E1 inhibitor PYR-41