Mov10 and APOBEC3G localization to processing bodies is not required for virion incorporation and antiviral activity.
Mov10 and APOBEC3G (A3G) localize to cytoplasmic granules called processing bodies (P bodies), incorporate into human immunodeficiency virus type 1 (HIV-1) virions, and inhibit viral replication. The functional relevance of Mov10/A3G P-body localization to virion incorporation and antiviral activity has not been fully explored. We found that a helicase V mutant of Mov10 exhibits significantly reduced localization to P bodies but still efficiently inhibits viral infectivity via virion incorporation. Disruption of the P bodies by DDX6 knockdown also confirmed Mov10 antiviral activity without P-body localization. In addition, overexpression of SRP19, which binds to 7SL RNA, depleted A3G from P bodies but did not affect its virion incorporation. Sucrose gradient sedimentation assays revealed that the majority of Mov10, A3G, HIV-1 RNA, and Gag formed high-molecular-mass (HMM) complexes that are converted to low-molecular-mass (LMM) complexes after RNase A treatment. In contrast, the P-body markers DCP2, LSM1, eIF4e, DDX6, and AGO1 were in LMM complexes, whereas AGO2, an effector protein of the RNA-induced silencing complex that localizes to P bodies, was present in both LMM and HMM complexes. Depletion of AGO2 indicated that RNA-induced silencing function is required for Mov10's ability to reduce Gag expression upon overexpression, but not its virion incorporation or effect on virus infectivity. We conclude that the majority of Mov10 and A3G are in HMM complexes, whereas most of the P-body markers are in LMM complexes, and that virion incorporation and the antiviral activities of Mov10 and A3G do not require their localization to P bodies.
Viral Mutation Section.
Taisuke
T
Izumi
Ryan
R
Burdick
Mayu
M
Shigemi
Sergey
S
Plisov
Wei-Shau
WS
Hu
Vinay K
VK
Pathak
D000071480:APOBEC-3G Deaminase; D000998:Antiviral Agents / Q000378:metabolism*; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D003564:Cytidine Deaminase / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism; D015497:HIV-1 / Q000737:chemistry / Q000276:immunology* / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D020365:RNA Helicases / Q000378:metabolism*; D014771:Virion / Q000737:chemistry / Q000276:immunology / Q000502:physiology; D019065:Virus Assembly
D000998:Antiviral Agents; C530476:Mov10 protein, human; D000071480:APOBEC-3G Deaminase; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase; D020365:RNA Helicases