Major autoantigenic sites of the (U1) small nuclear ribonucleoprotein-specific 68-kDa protein

A 68-kDa protein associated with (U1)snRNP is a major target for human autoantibodies to small ribonucleoprotein particles (snRNP) prevalent in a variety of inflammatory rheumatic diseases. The epitopes recognized by these antibodies were mapped by expression of subfragments of p68 cDNA in Escherichia coli and testing of the corresponding recombinant proteins for immunoreactivity with sera of patients with autoimmune diseases. Three of four antigenic regions were analysed in detail. The immunodominant autoantigenic region was found to coincide with the RNA-binding domain of the p68 protein and was shown to contain a nested set of overlapping discontinuous epitopes. Two additional non-overlapping major antigenic domains were localized in the carboxy-terminal half of the p68 protein. Each of these two carboxy-terminal domains was shown to contain more than one conformation-dependent epitope. Taking into account previous mapping studies, the data demonstrate that p68 contains at least four antigenic regions, each of which harbours multiple epitopes which are recognized in a patient-specific manner.