Open AccessA mammalian germ cell-specific RNA-binding protein interacts with ubiquitously expressed proteins involved in splice site selection
Author(s) -
David J. Elliott,
Cyril F. Bourgeois,
Albrecht Klink,
James Stévenin,
Howard J. Cooke
Publication year - 2000
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.97.11.5717
Subject(s) - rna splicing , biology , rna binding protein , alternative splicing , microbiology and biotechnology , sr protein , fusion protein , protein splicing , exonic splicing enhancer , genetics , rna , messenger rna , gene , recombinant dna
RNA-binding motif (RBM ) genes are found on all mammalian Y chromosomes and are implicated in spermatogenesis. Within human germ cells, RBM protein shows a similar nuclear distribution to components of the pre-mRNA splicing machinery. To address the function of RBM, we have used protein–protein interaction assays to test for possible physical interactions between these proteins. We find that RBM protein directly interacts with members of the SR family of splicing factors and, in addition, strongly interacts with itself. We have mapped the protein domains responsible for mediating these interactions and expressed the mouse RBM interaction region as a bacterial fusion protein. This fusion protein can pull-down several functionally active SR protein species from cell extracts. Depletion and add-back experiments indicate that these SR proteins are the only splicing factors bound by RBM which are required for the splicing of a panel of pre-mRNAs. Our results suggest that RBM protein is an evolutionarily conserved mammalian splicing regulator which operates as a germ cell-specific cofactor for more ubiquitously expressed pre-mRNA splicing activators.