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Temperature-sensitive mutation in yeast mitochondrial ribosome recycling factor (RRF)
Emeline Teyssier1, Go Hirokawa, Anna Tretiakova
1Department of Biochemistry and Molecular Pharmacology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107-5541, USA.
Nucleic Acids Research
|July 11, 2003
Summary
The yeast FIL1 gene product, Rrf1p, functions as a ribosome recycling factor (RRF) in mitochondria. Deleting RRF1 impairs mitochondrial protein synthesis, leading to respiratory deficiency and genome instability.
Area of Science:
- Mitochondrial biology
- Molecular genetics
- Protein synthesis
Background:
- The yeast FIL1 gene encodes Rrf1p, a protein homologous to Escherichia coli ribosome recycling factor (RRF).
- Rrf1p's cellular localization and specific function were previously uncharacterized.
Purpose of the Study:
- To investigate the function of yeast Rrf1p.
- To determine if Rrf1p acts as a ribosome recycling factor (RRF) in mitochondria.
Main Methods:
- Gene deletion and complementation in yeast (Saccharomyces cerevisiae).
- Mitochondrial genome stability assays (rho(-) petite analysis).
- Assessment of respiratory function on non-fermentable carbon sources.
- Measurement of mitochondrial and total protein synthesis rates at different temperatures.
Main Results:
- Rrf1p was localized to mitochondria.
- RRF1 deletion resulted in respiratory incompetence and mitochondrial genome instability.
- A temperature-sensitive rrf1 mutant exhibited severe inhibition of mitochondrial protein synthesis at the permissive temperature, leading to growth defects.
Conclusions:
- Yeast Rrf1p is essential for mitochondrial protein synthesis.
- Rrf1p functions as a ribosome recycling factor (RRF) within yeast mitochondria.