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PMID: 2951739 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 84 ·No. 8 ·1987-04-00 ·Pages 2421-5

Habets WJ, Sillekens PT, Hoet MH, Schalken JA, Roebroek AJ, Leunissen JA, van de Ven WJ, van Venrooij WJ

Abstract

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 us to isolate cDNA clone lambda HB''-1 from a phage lambda gt11 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 lambda HB''-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 lambda HB''-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 lambda HB''-1 cDNA insert and subsequent translation resulted in a protein product with the molecular size of the B'' protein. These data demonstrate that clone lambda HB''-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 biological function of autoimmune antigens.

MeSH Terms
Amino Acid Sequence Autoantigens/genetics Base Sequence Cloning, Molecular Connective Tissue Diseases/immunology DNA/analysis Humans Nucleic Acid Hybridization Peptide Mapping Protein Biosynthesis RNA, Small Nuclear/genetics Ribonucleoproteins/genetics,immunology Ribonucleoproteins, Small Nuclear
Chemicals
Autoantigens RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear DNA
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Habets W J
Sillekens P T
Hoet M H
Schalken J A
Roebroek A J
Leunissen J A
van de Ven W J
van Venrooij W J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.84.8.2421
Published
1987-04-00
Pages
2421-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC304663
Subset
IM
Databases
GENBANK
M15841
External Links
PubMed source DOI: 10.1073/pnas.84.8.2421 PMC Full Text View journal details
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