CDNA Cloning of a Human Autoimmune Nuclear Ribonucleoprotein Antigen

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1985 Dec 1

PMID 2999783

Citations 23

Authors

E D Wieben

A M Rohleder

J M Nenninger

T Pederson

Affiliations

Soon will be listed here.

Abstract

Sera from patients with systemic lupus erythematosus and other autoimmune disorders contain antibodies against nuclear proteins. One such autoantibody system, known as Sm, reacts with antigens associated with small nuclear RNA molecules. In this paper we report the use of Sm autoantibodies to isolate a cDNA clone for the mRNA of one of these nuclear antigens. A HeLa cell cDNA library was screened by message selection followed by autoantibody reaction of cell-free translation products. This led to the identification of a cDNA clone, p281, containing sequences complementary to mRNA for an Sm autoantibody-reactive, 11,000 Mr protein. This cloned Sm antigen comigrated with the small nuclear RNA-associated protein known as "E" and reacted with four out of four Sm autoantibodies that precipitate E protein from total mRNA translation products. RNA gel blot hybridization with clone p281 DNA revealed a poly(A)+ mRNA of approximately equal to 600 nucleotides in human and marmoset (New World primate) cells. Southern blot hybridization of HeLa cell and human lymphocyte DNA indicated the presence of 6-10 copies of p281-homologous sequences. Similar copy numbers were observed with genomic DNA from baboon, cat, and mouse, indicating that the Sm antigen mRNA sequence represented in p281 is conserved across three classes of the Mammalia (primates, carnivores, and rodents). However, no cross-hybridization of p281 was observed with frog or Drosophila DNA. In light of existing evidence that the mammalian Sm antigen E is a weaker autoantigen than other small nuclear RNA-associated proteins, these results suggest a possible correlation between a protein's capacity to serve as an autoantigen during breakdown of the host's immunological tolerance and its extent of evolutionary conservation, whereas the inverse relationship applies to conventional immunity. We suspect, as have others, that this is a clue to the mechanism of autoimmunity.

Citing Articles

Transcription of a variant human U6 small nuclear RNA gene is controlled by a novel, internal RNA polymerase III promoter.

Tichelaar J, Knerer B, Vrabel A, Wieben E Mol Cell Biol. 1994; 14(8):5450-7.

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Presence of antibodies to different subunits of replication protein A in autoimmune sera.

Garcia-Lozano R, Gonzalez-Escribano F, Sanchez-Roman J, Wichmann I, Nunez-Roldan A Proc Natl Acad Sci U S A. 1995; 92(11):5116-20.

PMID: 7761458 PMC: 41859. DOI: 10.1073/pnas.92.11.5116.

U1 precursors: variant 3' flanking sequences are transcribed in human cells.

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Identification of proteins that bind tightly to pre-mRNA during in vitro splicing.

Mayrand S, Pedersen N, Pederson T Proc Natl Acad Sci U S A. 1986; 83(11):3718-22.

PMID: 3459150 PMC: 323594. DOI: 10.1073/pnas.83.11.3718.

Loop I of U1 small nuclear RNA is the only essential RNA sequence for binding of specific U1 small nuclear ribonucleoprotein particle proteins.

Hamm J, van Santen V, Spritz R, Mattaj I Mol Cell Biol. 1988; 8(11):4787-91.

PMID: 2974920 PMC: 365571. DOI: 10.1128/mcb.8.11.4787-4791.1988.

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