title: Ro60 requires Y3 RNA for cell surface exposure and inflammation associated with cardiac manifestations of neonatal lupus.
abstract: Cardiac neonatal lupus (NL) is presumed to arise from maternal autoantibody targeting an intracellular ribonucleoprotein, Ro60, which binds noncoding Y RNA and only becomes accessible to autoantibodies during apoptosis. Despite the importance of Ro60 trafficking in the development of cardiac NL, the mechanism underlying cell surface exposure is unknown. To evaluate the influence of Y RNA on the subcellular location of Ro60 during apoptosis and activation of macrophages, stable Ro60 knockout murine fibroblasts expressing wild-type or mutated FLAG-Ro60 were assessed. FLAG3-Ro60(K170A R174A) binds Y RNA, whereas FLAG3-Ro60(H187S) does not bind Y RNA; fibroblasts expressing these constructs showed equivalent intracellular expression of Ro60. In contrast, apoptotic fibroblasts containing FLAG3-Ro60(K170A R174A) were bound by anti-Ro60, whereas FLAG3-Ro60(H187S) was not surface expressed. RNA interference of mY3 RNA in wild-type fibroblasts inhibited surface translocation of Ro60 during apoptosis, whereas depletion of mY1 RNA did not affect Ro60 exposure. Furthermore, Ro60 was not exposed following overexpression of mY1 in the mY3-depleted fibroblasts. In an in vitro model of anti-Ro60-mediated injury, Y RNA was shown to be an obligate factor for TLR-dependent activation of macrophages challenged with anti-Ro60-opsonized apoptotic fibroblasts. Murine Y3 RNA is a necessary factor to support the surface translocation of Ro60, which is pivotal to the formation of immune complexes on apoptotic cells and a TLR-dependent proinflammatory cascade. Accordingly, the Y3 RNA moiety of the Ro60 ribonucleoprotein imparts a critical role in the pathogenicity of maternal anti-Ro60 autoantibodies.
authors.affiliation: Department of Medicine, New York University School of Medicine, New York, NY 10016, USA. joanne.reed@nyumc.org
authors.forename: Joanne H
authors.initials: JH
authors.lastname: Reed
authors.forename: Soyeong
authors.initials: S
authors.lastname: Sim
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Wolin
authors.forename: Robert M
authors.initials: RM
authors.lastname: Clancy
authors.forename: Jill P
authors.initials: JP
authors.lastname: Buyon
mesh_terms: D000328:Adult; D000818:Animals; D000831:Animals, Newborn; D001323:Autoantibodies / Q000378:metabolism; D002478:Cells, Cultured; D002648:Child; D018920:Coculture Techniques; D006331:Heart Diseases / Q000276:immunology* / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000378:metabolism / Q000473:pathology; D008180:Lupus Erythematosus, Systemic / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D022661:RNA, Untranslated / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D013347:Subcellular Fractions / Q000276:immunology; D013499:Surface Properties; D000099207:SS-A Antigen
chemical_list: D001323:Autoantibodies; D022661:RNA, Untranslated; D012261:Ribonucleoproteins; D000099207:SS-A Antigen