title: Small nucleolar RNP scleroderma autoantigens associate with phosphorylated serine/arginine splicing factors during apoptosis.
abstract: OBJECTIVE
  Proteins that are phosphorylated during apoptosis are commonly precipitated by autoantibodies found in the sera of patients with systemic lupus erythematosus. We sought to determine whether scleroderma autoantigens such as small nucleolar RNPs (snoRNP) also associate with phosphoproteins in response to various cellular stressors.
  
  
  METHODS
  We screened a panel of monoclonal antibodies derived from mice exposed to mercury, a well-characterized murine model of the anti-snoRNP autoimmune response, for the ability to selectively precipitate phosphoproteins from radiolabeled lysates prepared from Jurkat T cells subjected to stressful stimuli.
  
  
  RESULTS
  Monoclonal antibodies reactive with snoRNPs precipitated a phosphoprotein complex (pp42, pp34, and pp23) from lysates prepared from apoptotic cells. Several novel phosphoproteins (pp62 and pp18) were also observed. The phosphorylation and/or recruitment of these proteins to the snoRNP complex is induced by multiple apoptotic stimuli (e.g., Fas ligation, anisomycin, or ultraviolet irradiation), an effect that is blocked by overexpression of Bcl-2. We were unable to demonstrate an association of the phosphoprotein complex with snoRNPs in cells treated with the xenobiotic agent mercury. The snoRNP-associated phosphoprotein complex is composed of serine/arginine (SR) splicing factors, including SRp40.
  
  
  CONCLUSIONS
  The association of phosphorylated SR proteins with snoRNPs in cells undergoing apoptosis suggests that the immune response to fibrillarin that characterizes a subset of patients with scleroderma may be related to cell death induced by apoptotic stimuli (e.g., Fas ligation, irradiation, or chemical toxins), or by exposure to mercury.
authors.affiliation: Denver Arthritis Clinic, Colorado, USA.
authors.forename: K
authors.initials: K
authors.lastname: Overzet
authors.forename: T J
authors.initials: TJ
authors.lastname: Gensler
authors.forename: S J
authors.initials: SJ
authors.lastname: Kim
authors.forename: M E
authors.initials: ME
authors.lastname: Geiger
authors.forename: W J
authors.initials: WJ
authors.lastname: van Venrooij
authors.forename: K M
authors.initials: KM
authors.lastname: Pollard
authors.forename: P
authors.initials: P
authors.lastname: Anderson
authors.forename: P J
authors.initials: PJ
authors.lastname: Utz
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001324:Autoantigens / Q000276:immunology*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells / Q000187:drug effects; D008628:Mercury / Q000494:pharmacology; D051379:Mice; D009687:Nuclear Proteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D010766:Phosphorylation / Q000528:radiation effects; D012326:RNA Splicing; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear / Q000276:immunology* / Q000378:metabolism; D012595:Scleroderma, Systemic / Q000276:immunology*; D000068103:Serine-Arginine Splicing Factors; D014466:Ultraviolet Rays
chemical_list: D001324:Autoantigens; D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear; C047458:fibrillarin; D000068103:Serine-Arginine Splicing Factors; D008628:Mercury