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Novel RNA-Binding Proteins Isolation by the RaPID Methodology
Published on: September 30, 2016
The Saccharomyces cerevisiae RNA-binding protein Rbp29 functions in cytoplasmic mRNA metabolism
E Winstall1, M Sadowski, U Kuhn
1Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.
The Journal of Biological Chemistry
|April 15, 2000
Summary
The Saccharomyces cerevisiae RBP29 gene encodes a cytoplasmic mRNA-binding protein. This protein interacts with poly(A) tail-binding protein and influences gene expression, particularly under stress conditions.
Area of Science:
- Molecular Biology
- Yeast Genetics
- RNA Biology
Background:
- The Saccharomyces cerevisiae RBP29 gene encodes a 29-kDa cytoplasmic protein, Rbp29p.
- Rbp29p is identified as an mRNA-binding protein involved in cytoplasmic mRNA regulation.
Purpose of the Study:
- To investigate the function and interactions of the RBP29 gene product in yeast.
- To determine the role of Rbp29p in mRNA processing and translation.
Main Methods:
- Co-immunoprecipitation assays to study protein interactions.
- In vitro binding assays using recombinant Rbp29p.
- Analysis of yeast strains with RBP29 deletion and overexpression.
Main Results:
- Rbp29p co-immunoprecipitates with poly(A) tail-binding protein Pab1p in a dosage- and RNA-dependent manner.
- Recombinant Rbp29p exhibits nanomolar binding affinity for poly(A) tails.
- RBP29 deletion exacerbates slow growth in yeast strains with eIF4G mutations (TIF4631, TIF4632).
- RBP29 overexpression suppresses temperature-sensitive growth defects in specific tif4631, tif4632, and pab1 alleles.
Conclusions:
- Rbp29p is an mRNA-binding protein that interacts with Pab1p and plays a role in modulating cytoplasmic mRNA expression.
- Rbp29p is not essential for viability but influences cellular growth and stress response pathways.
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