Isolation and partial characterization of messenger RNA, from murine T cell hybrids, coding for suppressive immunoglobulin G-binding factor.
Poly A RNA has been isolated from a murine T cell hybridoma ( T2D4 ) that spontaneously secretes suppressive immunoglobulin G-binding factor ( IgGBF ). Translation products, obtained from a rabbit reticulocyte lysate translation system and after injection into Xenopus laevis oocytes, contain material with the biologic activity, the affinity, and the m.w. of murine IgGBF ; it suppresses secondary in vitro IgG antibody production in a dose-dependent fashion. The suppressive factor binds to IgG but not to IgM immunoadsorbents and, after mild NaDodSO4 treatment, dissociates in NaDodSO4 polyacrylamide gels into two peaks at 78 and 40 kD. Translation products from two non- IgGBF -secreting cell lines (BW-5147, a T lymphoma line, and A9, a fibroblast cell line) fail to exert any suppressive activity. On sucrose gradients, the RNA responsible for the biologic activity was found in one major peak located at 11S. IgGBF synthesized in a cellfree translation system by using poly A RNA and sucrose gradient fractions was also characterized by immunoprecipitation with Fc fragments of [35S]methionine-labeled proteins. On NaDodSO4 polyacrylamide gels, it migrates in one peak located at 37 kD. We conclude that IgGBF is coded for by 11S poly A RNA and that no post-translational modifications (other than proteolytic cleavage) are necessary to obtain a biologically active factor with Ig-binding properties.
D000818:Animals; D000937:Antigen-Antibody Reactions; D002460:Cell Line; D011232:Chemical Precipitation; D005815:Genetic Code; D006825:Hybridomas / Q000276:immunology*; D007108:Immune Tolerance; D008222:Lymphokines / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008970:Molecular Weight; D029584:Prostatic Secretory Proteins; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000276:immunology / Q000302:isolation & purification*; D012156:Reticulocytes / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology
D008222:Lymphokines; D029584:Prostatic Secretory Proteins; D012333:RNA, Messenger; C043076:beta-microseminoprotein; C013059:immunoglobulin-binding factors