TY - JOUR
T1 - U3 snoRNP associates with fibrillarin a component of the scleroderma clumpy nucleolar domain
AU - Herrera-Esparza, Rafael
AU - Kruse, Lars
AU - von Essen, Marina
AU - Campos, Lourdes
AU - Barbosa, Olga
AU - Bollain, Juan-José
AU - Badillo, Isaías
AU - Avalos-Díaz, Esperanza
N1 - Keywords: Autoantigens; Base Sequence; Cell Nucleolus; Chromosomal Proteins, Non-Histone; DNA, Complementary; Enzyme-Linked Immunosorbent Assay; Gene Library; Hela Cells; Humans; Immune Sera; In Situ Hybridization; Molecular Sequence Data; Ribonucleoproteins, Small Nucleolar; Scleroderma, Systemic; Tissue Distribution
PY - 2002
Y1 - 2002
N2 - Serum from patients with scleroderma recognizes the clumpy autoantigen. The present studies addressed the issue as to whether the clumpy nucleolar autoantigen recognized by scleroderma serum is fibrillarin-U3 snoRNP. Clones encoding for clumpy autoantigen were immunodetected from a lambdagt11 HeLa cell random-primed library with the serum from a patient with diffuse scleroderma and autoautoantibodies against clumpy autoantigen. Sequences from the recombinant phages were amplified by PCR and subcloned into a pCRII vector. The DNA was sequenced by a dideoxy termination reaction. Ten lambdagt11 clumpy clones were detected by immunoscreening. One containing the glycine-rich and RNP2 fibrillarin domains was expressed in lysogenic bacteria. The recombinant proteins were used to elicit antibodies in rabbits, and these exhibited clumpy nucleolar reactivity. The recombinant fibrillarin tested by ELISA was recognized by the clumpy scleroderma serum from the majority of patients. In situ hybridization assays showed that the fibrillarin tagged by the elicited antibodies was colocalized with U3 snoRNP in the nucleolus in a clumpy manner and coprecipitated the U3 snoRNP. In conclusion, the fibrillarin-U3 snoRNP complex is the major component of the clumpy subcellular domain. Therefore these molecules constitute an important target of scleroderma autoantibodies.
AB - Serum from patients with scleroderma recognizes the clumpy autoantigen. The present studies addressed the issue as to whether the clumpy nucleolar autoantigen recognized by scleroderma serum is fibrillarin-U3 snoRNP. Clones encoding for clumpy autoantigen were immunodetected from a lambdagt11 HeLa cell random-primed library with the serum from a patient with diffuse scleroderma and autoautoantibodies against clumpy autoantigen. Sequences from the recombinant phages were amplified by PCR and subcloned into a pCRII vector. The DNA was sequenced by a dideoxy termination reaction. Ten lambdagt11 clumpy clones were detected by immunoscreening. One containing the glycine-rich and RNP2 fibrillarin domains was expressed in lysogenic bacteria. The recombinant proteins were used to elicit antibodies in rabbits, and these exhibited clumpy nucleolar reactivity. The recombinant fibrillarin tested by ELISA was recognized by the clumpy scleroderma serum from the majority of patients. In situ hybridization assays showed that the fibrillarin tagged by the elicited antibodies was colocalized with U3 snoRNP in the nucleolus in a clumpy manner and coprecipitated the U3 snoRNP. In conclusion, the fibrillarin-U3 snoRNP complex is the major component of the clumpy subcellular domain. Therefore these molecules constitute an important target of scleroderma autoantibodies.
U2 - 10.1007/s00403-002-0338-7
DO - 10.1007/s00403-002-0338-7
M3 - Journal article
C2 - 12373336
SN - 0340-3696
VL - 294
SP - 310
EP - 317
JO - Archives of Dermatological Research
JF - Archives of Dermatological Research
IS - 7
ER -