title: Innate immune signaling induces high levels of TC-specific deaminase activity in primary monocyte-derived cells through expression of APOBEC3A isoforms.
abstract: In HIV-1-infected individuals, G-to-A hypermutation is found in HIV-1 DNA isolated from peripheral blood mononuclear cells (PBMCs). These mutations are thought to result from editing by one or more host enzymes in the APOBEC3 (A3) family of cytidine deaminases, which act on CC (APOBEC3G) and TC (other A3 proteins) dinucleotide motifs in DNA (edited cytidine underlined). Although many A3 proteins display high levels of deaminase activity in model systems, only low levels of A3 deaminase activity have been found in primary cells examined to date. In contrast, here we report high levels of deaminase activity at TC motifs when whole PBMCs or isolated primary monocyte-derived cells were treated with interferon-alpha (IFNalpha) or IFNalpha-inducing toll-like receptor ligands. Induction of TC-specific deaminase activity required new transcription and translation and correlated with the appearance of two APOBEC3A (A3A) isoforms. Knockdown of A3A in monocytes with siRNA abolished TC-specific deaminase activity, confirming that A3A isoforms are responsible for all TC-specific deaminase activity observed. Both A3A isoforms appear to be enzymatically active; moreover, our mutational studies raise the possibility that the smaller isoform results from internal translational initiation. In contrast to the high levels of TC-specific activity observed in IFNalpha-treated monocytes, CC-specific activity remained low in PBMCs, suggesting that A3G deaminase activity is relatively inhibited, unlike that of A3A. Together, these findings suggest that deaminase activity of A3A isoforms in monocytes and macrophages may play an important role in host defense against viruses.
authors.affiliation: Department of Global Health, University of Washington, Seattle, Washington 98102, USA.
authors.forename: Beth K
authors.initials: BK
authors.lastname: Thielen
authors.forename: John P
authors.initials: JP
authors.lastname: McNevin
authors.forename: M Juliana
authors.initials: MJ
authors.lastname: McElrath
authors.forename: Brook Vander Stoep
authors.initials: BV
authors.lastname: Hunt
authors.forename: Kevin C
authors.initials: KC
authors.lastname: Klein
authors.forename: Jaisri R
authors.initials: JR
authors.lastname: Lingappa
mesh_terms: D000071480:APOBEC-3G Deaminase; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002460:Cell Line; D003564:Cytidine Deaminase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic / Q000276:immunology*; D055785:Gene Knockdown Techniques; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics*; D016898:Interferon-alpha / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000201:enzymology / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000201:enzymology* / Q000276:immunology* / Q000378:metabolism; D014176:Protein Biosynthesis / Q000276:immunology; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015398:Signal Transduction / Q000235:genetics*; D013379:Substrate Specificity; D051193:Toll-Like Receptors / Q000378:metabolism; D014158:Transcription, Genetic / Q000276:immunology
chemical_list: D016898:Interferon-alpha; D011506:Proteins; D034741:RNA, Small Interfering; D051193:Toll-Like Receptors; D000071480:APOBEC-3G Deaminase; C121399:APOBEC3A protein, human; C464924:APOBEC3G protein, human; D003564:Cytidine Deaminase