title: Endogenous APOBEC3A DNA cytosine deaminase is cytoplasmic and nongenotoxic.
abstract: APOBEC3A (A3A) is a myeloid lineage-specific DNA cytosine deaminase with a role in innate immunity to foreign DNA. Previous studies have shown that heterologously expressed A3A is genotoxic, suggesting that monocytes may have a mechanism to regulate this enzyme. Indeed, we observed no significant cytotoxicity when interferon was used to induce the expression of endogenous A3A in CD14(+)-enriched primary cells or the monocytic cell line THP-1. In contrast, doxycycline-induced A3A in HEK293 cells caused major cytotoxicity at protein levels lower than those observed when CD14(+) cells were stimulated with interferon. Immunofluorescent microscopy of interferon-stimulated CD14(+) and THP-1 cells revealed that endogenous A3A is cytoplasmic, in stark contrast to stably or transiently transfected A3A, which has a cell-wide localization. A3A constructs engineered to be cytoplasmic are also nontoxic in HEK293 cells. These data combine to suggest that monocytic cells use a cytoplasmic retention mechanism to control A3A and avert genotoxicity during innate immune responses.
authors: Department of Biochemistry, Molecular Biology and Biophysics, Institute for Molecular Virology, Masonic Cancer Center, and Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455, USA.
authors: Allison M
authors: AM
authors: Land
authors: Emily K
authors: EK
authors: Law
authors: Michael A
authors: MA
authors: Carpenter
authors: Lela
authors: L
authors: Lackey
authors: William L
authors: WL
authors: Brown
authors: Reuben S
authors: RS
authors: Harris
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival; D003564:Cytidine Deaminase / Q000502:physiology*; D003593:Cytoplasm / Q000201:enzymology*; D004249:DNA Damage; D015870:Gene Expression; D057809:HEK293 Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D007113:Immunity, Innate; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D009000:Monocytes / Q000201:enzymology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D011506:Proteins / Q000502:physiology*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C468783:H2AX protein, human; D006657:Histones; D018950:Lipopolysaccharide Receptors; D011506:Proteins; D012097:Repressor Proteins; C474831:TRIB3 protein, human; D017346:Protein Serine-Threonine Kinases; C121399:APOBEC3A protein, human; D003564:Cytidine Deaminase
keywords: APOBEC3A; Cytoplasmic Localization; DNA Cytosine Deamination; DNA Damage; Gamma-H2AX; Genotoxicity; HIV; Innate Immunity; Interferon; Monocytes
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