Abstract
In characterizing a series of yeast (Saccharomyces cerevisiae) mutants synthetic lethal with U1 RNA, we have identified a yeast gene (MUD2) with sequence similarity to the well-studied metazoan splicing factor U2AF65. The biochemical characterization indicates that the MUD2 gene product (MUD2P) contacts pre-mRNA directly and is a component of the pre-mRNA-U1 snRNP complex (commitment complex) that forms during early spliceosome assembly in yeast extracts. Unlike U1 snRNP itself, the association of MUD2P with pre-mRNA is dependent on a proper yeast branchpoint sequence. Genetic experiments indicate that MUD2P affects U2 snRNP addition. Moreover, experiments in the two-hybrid system show that PRP11P, a recently identified component of U2 snRNP, can interact directly with MUD2P. The experiments identify a specific inter-snRNP protein-protein contact that occurs during spliceosome assembly and more generally support substantial functional similarity between U2AF65 and MUD2P.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Amino Acid Sequence
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Cloning, Molecular
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Consensus Sequence
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Fungal Proteins / chemistry
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Fungal Proteins / genetics
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Fungal Proteins / metabolism*
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Genes, Lethal
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Hemagglutinin Glycoproteins, Influenza Virus
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Hemagglutinins, Viral / metabolism
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Models, Genetic
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Molecular Sequence Data
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RNA Precursors / metabolism*
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RNA Processing, Post-Transcriptional
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RNA-Binding Proteins
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Recombinant Fusion Proteins / biosynthesis
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Ribonucleoprotein, U1 Small Nuclear / metabolism
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Ribonucleoprotein, U2 Small Nuclear / metabolism*
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Ribonucleoproteins / chemistry
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Ribonucleoproteins / genetics
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Ribonucleoproteins / metabolism*
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae Proteins*
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Sequence Alignment
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Spliceosomes / metabolism
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Splicing Factor U2AF
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Viral Envelope Proteins / metabolism
Substances
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Fungal Proteins
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Hemagglutinin Glycoproteins, Influenza Virus
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Hemagglutinins, Viral
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MUD2 protein, S cerevisiae
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PRP11 protein, S cerevisiae
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RNA Precursors
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RNA-Binding Proteins
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Recombinant Fusion Proteins
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Ribonucleoprotein, U1 Small Nuclear
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Ribonucleoprotein, U2 Small Nuclear
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Ribonucleoproteins
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Saccharomyces cerevisiae Proteins
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Splicing Factor U2AF
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Viral Envelope Proteins