Can yeast be used to study mitochondrial diseases? Biolistic tRNA mutants for the analysis of mechanisms and suppressors.
Base substitutions equivalent to those causing human pathologies have been introduced in yeast mitochondrial tRNA genes. These mutants can be utilized as flexible tools to investigate the molecular aspects of mitochondrial diseases and identify correcting genes. We show that for all studied tRNA mutations (including an homoplasmic one in tRNA(Val)) the severity of phenotypes follows the same trend in four different nuclear backgrounds. Correcting genes include TUF1 and genes encoding aminoacyl-tRNA synthetase. The effect of suppressors was analyzed by Northern blot. Mutated leucyl-tRNA synthetase with highly reduced catalytic activity maintains full suppressing effect, thus suggesting a chaperone-like and/or stabilizing function.
De Luca
Cristina
C
Department of Cell and Developmental Biology, Pasteur Institute-Fondazione Cenci Bolognetti, University Sapienza, Rome, Italy.
Zhou
YouFang
Y
Montanari
Arianna
A
Morea
Veronica
V
Oliva
Romina
R
Besagni
Celine
C
Bolotin-Fukuhara
Monique
M
Frontali
Laura
L
Francisci
Silvia
S
D001483:Base Sequence; D019470:Biolistics / Q000379:methods*; D015152:Blotting, Northern; D006801:Humans; D050296:Microbial Viability; D028361:Mitochondrial Diseases / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010641:Phenotype; D017354:Point Mutation; D012343:RNA, Transfer / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D013489:Suppression, Genetic
D029701:Saccharomyces cerevisiae Proteins; D012343:RNA, Transfer