Monoclonal antibody specific to a subclass of polyproline-Arg motif provides evidence for the presence of an snRNA-free spliceosomal Sm protein complex in vivo: implications for molecular interactions involving proline-rich sequences of Sm B/B' proteins.
The human spliceosomal Sm B/B' proteins are essential for the biogenesis of the snRNP particles. B/B' proteins contain several clusters of the PPPPGM/IR sequence, which occurs within the C-terminus of Sm B/B'. This sequence is very similar to the PPPPPGHR sequence of the cytoplasmic tail of the CD2 receptor and closely resembles the class II of SH3 ligands, suggesting a similarly important role. We report that a monoclonal antibody (3E10) against the PPPPPGHR sequence recognizes spliceosomal Sm B/B' proteins. Proteins that are specifically immunoprecipitated by 3E10 include Sm B, B', D1, D2, D3, E, F, and G. However, unlike Y12 and other anti-Sm immunoprecipitates, 3E10 immunoprecipitates appear to lack the U1 snRNP-specific proteins A and C and U snRNAs. These findings indicate that 3E10 recognizes a subset of Sm protein core and suggest the presence of snRNA-free Sm protein complex(es) in vivo. We propose that the epitope binding for 3E10 may become unaccessible upon interactions of Sm proteins and their subsequent incorporation into the core particles. The Sm proline-rich sequences may have an important role in mediating protein-protein interactions necessary for the proper snRNP core assembly or function, or both. To our knowledge, 3E10 is the first well characterized mAb specific for a subclass of polyproline-arg motif recognizing Sm B/B' and CD2 proteins. 3E10 antibody can be used to further characterize the nature of protein components in the snRNA-free Sm subcore protein complex(es) that are formed during the snRNP core assembly steps.
Department of Pathology, University of Pennsylvania, School of Medicine, Philadelphia 19104-4283, USA.
M
M
Filali
J
J
Qiu
S
S
Awasthi
U
U
Fischer
D
D
Monos
M
M
Kamoun
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D001120:Arginine / Q000276:immunology; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004151:Dipeptides / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011233:Precipitin Tests; D012342:RNA, Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D017381:Spliceosomes / Q000378:metabolism; D055517:snRNP Core Proteins
D000911:Antibodies, Monoclonal; D001324:Autoantigens; D004151:Dipeptides; D000939:Epitopes; D010455:Peptides; D012342:RNA, Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear; D055517:snRNP Core Proteins; C011083:polyproline; D001120:Arginine