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Title: Analysis of a cDNA clone expressing a human autoimmune antigen: full-length sequence of the U2 small nuclear RNA-associated B antigen

A U2 small nuclear RNA-associated protein, designated B'', was recently identified as the target antigen for autoimmune sera from certain patients with systemic lupus erythematosus and other rheumatic diseases. Such antibodies enabled them to isolate cDNA clone lambdaHB''-1 from a phage lambdagt11 expression library. This clone appeared to code for the B'' protein as established by in vitro translation of hybrid-selected mRNA. The identity of clone lambdaHB''-1 was further confirmed by partial peptide mapping and analysis of the reactivity of the recombinant antigen with monospecific and monoclonal antibodies. Analysis of the nucleotide sequence of the 1015-base-pair cDNA insert of clone lambdaHB''-1 revealed a large open reading frame of 800 nucleotides containing the coding sequence for a polypeptide of 25,457 daltons. In vitro transcription of the lambdaHB''-1 cDNA insert and subsequent translation resulted in a protein product with the molecular size of the B'' protein. These data demonstrate that clone lambdaHB''-1 contains the complete coding sequence of this antigen. The deduced polypeptide sequence contains three very hydrophilic regions that might constitute RNA binding sites and/or antigenic determinants. These findings might have implications both for the understanding of the pathogenesis of rheumatic diseases as well as for the elucidation of the biologicalmore » function of autoimmune antigens.« less
Authors:
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Publication Date:
OSTI Identifier:
6077626
Resource Type:
Journal Article
Resource Relation:
Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States); Journal Volume: 84:8
Research Org:
Univ. of Nijmegen, Netherlands
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ANTIGENS; AMINO ACID SEQUENCE; DNA SEQUENCING; RHEUMATIC DISEASES; PATHOGENESIS; SULFUR 35; TRACER TECHNIQUES; HELA CELLS; IMMUNE SERUMS; IN VITRO; METHIONINE; PATIENTS; PROTEINS; RECOMBINANT DNA; RNA; TRANSCRIPTION; AMINO ACIDS; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; CARBOXYLIC ACIDS; DAYS LIVING RADIOISOTOPES; DISEASES; DNA; DRUGS; EVEN-ODD NUCLEI; ISOTOPE APPLICATIONS; ISOTOPES; LIGHT NUCLEI; LIPOTROPIC FACTORS; MOLECULAR STRUCTURE; NUCLEI; NUCLEIC ACIDS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; RADIOISOTOPES; SKELETAL DISEASES; STRUCTURAL CHEMICAL ANALYSIS; SULFUR ISOTOPES 550201* -- Biochemistry-- Tracer Techniques