Autoimmunity to RNA polymerase II is focused at the carboxyl terminal domain of the large subunit

Michito Hirakata, Jyotshna Kanungo, Akira Suwa, Yoshihiko Takeda, Joe Craft, John A. Hardin

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Objective. Previous studies have demonstrated antibodies to the large (220 kd) polypeptide subunit of RNA polymerase II (Pol II) in sera from certain patients with scleroderma. In the present study, we sought to identify the autoantigenic region on this polypeptide. Methods. A recombinant fusion protein, corresponding to the 52-heptapeptide repeat found in the carboxyl terminal domain (CTD) of the large Pol II subunit, was used to identify 15 patient sero that contained autoantibodies. Synthetic peptides CTD7 (representing a single heptapeptide) and CTD18 (representing 2 1/4 heptapeptide repeats) were used in a competitive inhibition assay to define the specificity of these sero and the importance of the CTD as an autoantigen. Results. All 15 sera immunoprecipitated the Pol II subunit from radiolabeled cell extracts, and 11 of them bound the CTD fusion protein in immunoblots. Immunoprecipitation of Pol II was completely inhibited by CTD18 in 5 sera and partially inhibited in 4 additional sero. Conclusion. These results indicate that the CTD heptapeptide repeat is a focal point for autoimmune responses in scleroderma. It is likely that the repetitive sequence and high content of charged residues of this structure contribute to its role as an autoantigen.

Original languageEnglish
Pages (from-to)1886-1891
Number of pages6
JournalArthritis and Rheumatism
Volume39
Issue number11
DOIs
Publication statusPublished - 1996 Nov

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RNA Polymerase II
Autoimmunity
Autoantigens
Peptides
Serum
Recombinant Fusion Proteins
Nucleic Acid Repetitive Sequences
Cell Extracts
Immunoprecipitation
Autoantibodies
Antibodies

ASJC Scopus subject areas

  • Immunology
  • Rheumatology

Cite this

Autoimmunity to RNA polymerase II is focused at the carboxyl terminal domain of the large subunit. / Hirakata, Michito; Kanungo, Jyotshna; Suwa, Akira; Takeda, Yoshihiko; Craft, Joe; Hardin, John A.

In: Arthritis and Rheumatism, Vol. 39, No. 11, 11.1996, p. 1886-1891.

Research output: Contribution to journalArticle

Hirakata, Michito ; Kanungo, Jyotshna ; Suwa, Akira ; Takeda, Yoshihiko ; Craft, Joe ; Hardin, John A. / Autoimmunity to RNA polymerase II is focused at the carboxyl terminal domain of the large subunit. In: Arthritis and Rheumatism. 1996 ; Vol. 39, No. 11. pp. 1886-1891.
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