Multiple protein: protein interactions between the snRNP common core proteins.

Abstract

The snRNP core proteins (B, D3, D2, D1, E, F, and G) assemble with snRNA and form the snRNP core particle with a suggested stoichiometry of B2[D1, D2(E, F, G)2]D3. The newly synthesized snRNP core proteins are stored in the cytoplasm in three RNA-free complexes of (1) B at 2S-6S; (2) [D1, D2(E, F, G)2] at 6S; and (3) (B, D3, and 69 kDa) at 20S. The snRNP proteins assemble stepwise with snRNAs that appear transiently in the cytoplasm before returning to the nucleus as mature snRNP particles. In this report, two approaches are used to investigate the protein:protein interactions between the snRNP proteins. First, the 6S and 20S cytoplasmic complexes chromatographed as intact structures, supporting their identifications as discrete complexes. Second, the cDNAs for the proteins were used to test all pair-wise interactions between the seven major core proteins using the yeast two-hybrid system. The two-hybrid system identified four strong reciprocal interactions, one weak reciprocal interaction, five one-way interactions, and one homotypic interaction. The strongest interactions were between proteins within the 6S particle. Other interactions were between proteins in the 6S and 20S particles or within the 20S particle itself. These interactions are likely to occur within the cytoplasmic snRNP core protein complexes and the mature snRNP particle.