Abstract
We previously reported the isolation of PRP28, a gene in Saccharomyces cerevisiae whose activity is required for the first step of nuclear mRNA splicing in vivo. Sequence analysis revealed that PRP28 is included in the 'DEAD-box' gene family, members of which are thought to function as ATP-dependent RNA helicases. Genetic interactions led us to suggest that PRP28 is functionally associated with the U4/U5/U6 snRNP. We have now purified the PRP28 protein from S. cerevisiae and demonstrated that it is required for the first step of splicing in vitro. Interestingly, PRP28 is not a stably associated snRNP protein. Strand displacement assays indicate that PRP28 does not exhibit RNA helicase activity, suggesting that an additional factor or factors may be required for its activation.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / pharmacology
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Anthranilate Synthase / genetics
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Antibodies / pharmacology
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DEAD-box RNA Helicases
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Escherichia coli / genetics
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RNA / pharmacology
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RNA Helicases
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RNA Nucleotidyltransferases / genetics
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RNA Nucleotidyltransferases / isolation & purification
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RNA Nucleotidyltransferases / metabolism*
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RNA Splicing*
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RNA, Messenger / metabolism*
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae / chemistry*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae Proteins
Substances
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Antibodies
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RNA, Messenger
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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RNA
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Adenosine Triphosphate
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RNA Nucleotidyltransferases
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Adenosine Triphosphatases
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PRP28 protein, S cerevisiae
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DEAD-box RNA Helicases
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RNA Helicases
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Anthranilate Synthase