Mitochondrial release factor in yeast: interplay of functional domains.
The MRF1 gene encodes the only class I release factor found in Saccharomyces cerevisiae mitochondria, mRF1. The previously isolated point mutation mrf1-13 caused respiratory deficiency due to inhibition of mitochondrial translation. In this study, we have isolated second-site suppressors of mrf1-13. Among over 200 respiratory positive suppressor colonies, ten nuclear dominant suppressors had a new mutation in the MRF1 gene. The suppressors in combination with the original mrf1-13 revealed increased levels of mitochondrially synthesized proteins, Cox2 and Atp6. One of the suppressor alleles was cloned on a plasmid and was found to support weaker respiratory competence than in combination with mrf1-13. Finally, the possible effects of the suppressor mutations are discussed based on a structural model of mRF1 protein built for its "open" and "closed" forms using known crystal structures of prokaryotic release factor RF1 as templates. The 3D models suggest that at least some suppressors switch the structure of mRF1 from the "closed" to a permanently "open" form causing stronger binding of the mRF1 protein to the ribosome and increasing the time of ribosome occupation. This explains how the suppressor mutants may facilitate translation termination despite a defect in decoding of the stop signal.
Kutner
Jan
J
Department of Genetics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Pawinskiego 5a, 02-106, Warsaw, Poland.
Towpik
Joanna
J
Ginalski
Krzysztof
K
Boguta
Magdalena
M
D000483:Alleles; D015153:Blotting, Western; D015966:Gene Expression Regulation, Fungal; D008928:Mitochondria; D024101:Mitochondrial Proteins; D010454:Peptide Termination Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012270:Ribosomes / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D013489:Suppression, Genetic / Q000235:genetics*
C507208:MRF1 protein, S cerevisiae; D024101:Mitochondrial Proteins; D010454:Peptide Termination Factors; D029701:Saccharomyces cerevisiae Proteins