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Binding of a cell-type-specific RNA splicing factor to its target regulatory sequence
1Department of Biology, Yale University, New Haven, Connecticut 06520-8103.
Molecular and Cellular Biology
|April 1, 1995
Summary
The Mer1 protein regulates splicing of the Saccharomyces cerevisiae MER2 gene during meiosis. This RNA-binding protein specifically interacts with MER2 RNA, enhancing its splicing efficiency.
Area of Science:
- Molecular Biology
- Yeast Genetics
- RNA Splicing
Background:
- The MER2 gene transcript in Saccharomyces cerevisiae undergoes efficient splicing during meiosis but not vegetative growth.
- Splicing of the MER2 transcript requires the Mer1 protein, which is meiosis-specific.
- The 5' exon of MER2 is unusually large and plays a role in splicing regulation.
Purpose of the Study:
- To investigate the role of the Mer1 protein in MER2 gene splicing.
- To identify cis-acting sequences involved in Mer1-dependent splicing.
- To determine the mechanism by which Mer1 regulates MER2 splicing.
Main Methods:
- Analysis of deletion and substitution mutations in the MER2 gene.
- Construction and analysis of MER2 deletion mutants and hybrid genes.
- RNA gel mobility shift assays and RNase T1 footprinting to study Mer1-RNA interactions.
Main Results:
- The 80-base MER2 intron is sufficient for Mer1-dependent splicing in vivo; 5' exon sequences enhance efficiency.
- Mer1 protein binds specifically to MER2 RNA, with highest affinity for the intact substrate.
- Mer1 protein contacts both the 5' exon and intron sequences of MER2 RNA.
Conclusions:
- Mer1 protein directly interacts with regulatory elements in MER2 RNA.
- Mer1 protein binding to MER2 RNA promotes its splicing during meiosis.
- The Mer1 protein acts as a specific regulator of MER2 splicing in Saccharomyces cerevisiae.