At least three linear regions but not the zinc-finger domain of U1C protein are exposed at the surface of the protein in solution and on the human spliceosomal U1 snRNP particle

No structural information on U1C protein either in its free state or bound to the spliceosomal U1 small nuclear ribonucleoprotein (snRNP) particle is currently available. Using rabbit antibodies raised against a complete set of 15 U1C overlapping synthetic peptides (16-30 residues long) in different immunochemical tests, linear regions exposed at the surface of free and U1 . snRNP-bound U1C were identified. Epitopes within at least three regions s panning residues 31-62, 85-103 and 116-159 were recognized on free and plas tic-immobilized recombinant human U1C expressed in Escherichia coil, on in vitro translated U1C protein and on U1C bound to the U1 snRNP particle pres ent in HeLa S100 extract. Using a zinc affinity labeling method, we further showed that the N-terminal U1C peptide containing a zinc-finger motif (pep tide 5-34) effectively binds Zn-65(2+). The N-terminal region of U1C, which is functional in U1 snRNP assembly, is apparently not located at the surfa ce of the U1 snRNP particle.