Identification and characterization of a calreticulin-binding nuclear protein as histone (H1),an autoantigen in systemic lupus erythematosus

Abstract

Our objective was to identify nuclear calreticulin-binding protein(s) and investigate whether there is a correlation between presence of autoantibodies against calreticulin and calreticulin-binding protein(s) in the sera of patients suffering from systemic lupus erythematosus (SLE).

The ligand overlay procedure using digoxigenin-labelled calreticulin was used to identify a calreticulin-binding protein in the nuclear fraction of bovine brain. Fractionation of the nuclear components was used to localize the major positive calreticulin-binding protein. The protein was partially purified using hydroxylapatitie chromatography and subjected to NH²-amino acid sequence analysis. Immunoblots using the sera of SLE patients were then carried out on calreticulin and the calreticulin-binding protein.

The calreticulin-binding protein present in the nucleoplasm was identified as histone H1. Approximately 62% (26=42) patients with SLE had IgG antibodies directed against H1 whereas the sera of healthy individuals did not react with the antigen; 36% of patients with SLE had both anticalreticulin and anti-histone H1 antibodies. Phosphorylation of the latter protein did not alter its immunoreactivity.

These findings demonstrate that the concomitant presence of autoantibodies directed against both calreticulin and histone H1 occurs frequently in patients with SLE and may help shed some light on the mechanisms which bring about the autoimmune response.

Keywords

calreticulin histone 1 systemic lupus erythematosus autoantibodies

Get full access to this article

View all access options for this article.

References

Tan EM . Antinuclear antibodies: a diagnostic marker for autoimmune disease and probes for cell biology . Adv Immunol 1989; 44: 93–151 .

Woods VL Jr . Systemic lupus erythematosus and related syndrome. In: Kelley WN , Harris ED , Ruddy S , Sledge CB (eds) Rheumatology WB Saunders Company: Philapdelphia , 1993.

Isenberg DA , Horsfall AC . Systemic lupus erythematosus—adult onset. In: Maddison PJ , Isenberg DA , Woo P , Glass DN (eds) Textbook of Rheumatology. Oxford University Press Inc: New York , 1993.

Villarreal GM et al. Prevalence of 13 autoantibodies and of the 16=6 and related pathogenic idiotypes in 465 patients with systemic lupus erythematosus and their relationship with disease activity . Lupus 1997; 6: 425–435 .

Lefkowith JB , Gilkenson GS . Nephritogenic autoantibodies in lupus . Arthritis Rheum 1996; 39: 849–903 .

Spronk PE et al. Levels of soluble VCAM-1, soluble ICAM-1 and soluble E-selectin during disease exacerbations in patients with systemic lupus erythematosus (SLE); a long term prospective study . Clin Exp Immunol 1994; 97: 439–444 .

Pisetsky DS . Antinuclear antibodies . Rheum Dis Clin North Am 1992; 18: 415–437 .

Vianna JL et al. Comparison of the primary and secondary antiphospholipid syndrome: a European multicenter study of 114 patients . Am J Med 1994; 96: 3–9 .

Isenberg DA et al. The relationship of anti-DNA antibodies idiotypes and cardiolipin antibodies to disease activity in systemic lupus erythematosus . Medicine 1986; 65: 46–55 .

10.

Rokeach LA et al Characterization of the autoantigen calreticulin . J Immunol 1991; 147: 3031–3039 .

11.

Michalak M , Milner RE , Bums K , Opas M . Calreticulin . Biochem J 1992; 285: 681–682 .

12.

Spiro RG , Zhu Q , Bhoyroo V , Soling HD . Definition of the lectin-like properties of the molecular chaperone, calreticulin, and demonstration of its copurification with endomannosidase from rat liver Golgi . J Biol Chem 1996; 271: 11588–11594 .

13.

Dupuis M , Schaerer E , Krause KH , Tschopp J . The calcium-binding protein calreticulin is a major constituent of lytic granules in cytolytic T-lymphocytes . J Exp Med 1993; 717: 1–7 .

14.

White TK , Zhu Q , Tanzer ML . Cell surface calreticulin is a putative mannoside lectin which triggers mouse melanoma cell spreading . J Biol Chem 1995; 270: 15926–15929 .

15.

Burns K et al. Modulation of gene expression by calreticulin binding to the glucocorticoid receptor . Nature 1994; 367: 476–480 .

16.

Tan EM , Cohen AS , Fries JF . The 1982 revised criteria for the classification of systemic lupus erythematosus . Arthritis Rheum 1982; 25: 1271–1277 .

17.

Treves S et al. Calreticulin is a candidate for a calsequestrin-like function in a Ca2/—storage compartments (calciosomes) of liver and brain . Biochem J 1990; 712: 473–480 .

18.

Tata JR . Isolation of nuclei from liver and other tissues . Methods Enzymol 1974; 31: 253–262 .

19.

Gurr MI , Fincan JB , Hawthome JN . The phospholipids of liver-cell fractions . Biochem Biophys Acta 1963; 70: 406–416 .

20.

Gilchrist JSC , Pierce GN . Identification and purification of a calcium binding protein in hepatic nuclear membranes . J Biol Chem 1993; 268: 4291–4299 .

21.

Bradford MM . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding . Anal Biochem 1976; 72: 248–254 .

22.

Laemmli UK . Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature 1970; 227: 680–685 .

23.

Guerrini R et al. Calmodulin binding sites of the skeletal, cardiac and brain ryanodine receptor Ca2/ channels: modulation by the catalytic subunit of cAMP-dependent protein kinase? Biochemistry 1995; 34: 5120–5129 .

24.

Aarden LA et al. Detection of antibodies to DNA by radioimmunoassay and immunofluorescence . Scand J Rheumatol 1975; (Suppl) 11: 12–19 .

25.

Roth SY , Allis CD . Chromatin decondensation: does histone H1 dephosphorylation play a role? Trends Biochem Sci 1992; 17: 93–98 .

26.

Singh NK , Atreya CD , Hakhashi HL . Identification of calreticulin as a rubella virus RNA binding protein . Proc Natl Acad Sci USA 1994; 91: 12770–12774 .

27.

Cheng ST et al. Calreticulin binds HYRNA and the 2 kDa polypeptide component of the Ro/SS-A ribonucleoprotein autoantigen . J Immunol 1996; 156: 4484–4491 .

28.

Wheeler DG et al. Calreticulin inhibits vitamin D3 signal transduction . Nucleic Acids Res 1995; 23: 3268–3274 .

29.

Dedhar S et al. Inhibition of nuclear hormone receptor activity by calreticulin . Nature 1994; 367: 480–483 .

30.

Burns K , Michalak M . Interactions of calreticulin with proteins of the endoplasmic and sarcoplasmic reticulum membranes . FEBS Lett 1993; 318: 181–185 .

31.

Eggleton P , Reid KBM , Kishore U , Sontheimer RD . Clinical relevance of calreticulin in systemic lupus erythematosus . Lupus 1997; 6: 564–571 .

32.

Lieu TS , Newkirk MM , Capra JD , Sonthelmer RD . Molecular characterization of human Ro/SS-A antigen. Amino terminal sequence of the protein moiety of human Ro/SS-A antigen and immunological activity of a corresponding synthetic peptide . J Clin Invest 1988; 82: 96–101 .

33.

Gioud M , Kaci MA , Monier JC . Histone antibodies in systemic lupus erythematosus . Arthritis Rheum 1982; 25: 407–413 .

34.

Fritzler MJ . Histone antibodies. In: DJ Wallace , BH Hahn (eds) ‘Dubois’ lupus erythematosus. Lee & Febinger: Pennsylvania , 1993.

35.

Stemmer CI , Muller S . Histone autoantibodies other than (H2A-H2B0DNA autoantibodies. In: Peter JB , Shoenfeld Y (eds) Autoantibodies. Elsevier Science, B.V. , 1996.

36.

von Muhlen CA , Tan EM . Autoantibodies in the diagnosis of systemic rheumatic diseases . Sem Arthritis Rheum 1995; 24: 323–358 .

37.

Tseng C-E et al. The 52-kd protein as a target of intermolecular spreading of the immune response to components of the SS-A/R–SSB/La complex . Arthritis Rheum 1997; 40: 936–944 .

38.

Bruckner RC , Cox MM . The histone-like H protein of E coli is ribosomal protein S3 . Nucleic Acids Res 1989; 17: 3145–3161 .

39.

McCauliffe DP et al. A human Ro/SS-A autoantigen is the homologue of calreticulin and is highly homologous with onchoncercal RAL-1 antigen and an aplysia ‘memory molecule’ . J Clin Invest 1990; 86: 332–335 .

40.

Khalife J et al. Cloning of the gene encoding a Schistosoma mansoni antigen homologus to human Ro/SSA autoantigen . Mol Biochem Parasitol 1993; 57: 193–202 .

41.

Ohe Y , Hayashi H , Iwai K . Human spleen histone-H1: isolation and amino acid sequences of three minor variants, H1a; Hic and Hid . JH Biochem 1989; 106: 844–857 .