Computationally driven, quantitative experiments discover genes required for mitochondrial biogenesis.
Mitochondria are central to many cellular processes including respiration, ion homeostasis, and apoptosis. Using computational predictions combined with traditional quantitative experiments, we have identified 100 proteins whose deficiency alters mitochondrial biogenesis and inheritance in Saccharomyces cerevisiae. In addition, we used computational predictions to perform targeted double-mutant analysis detecting another nine genes with synthetic defects in mitochondrial biogenesis. This represents an increase of about 25% over previously known participants. Nearly half of these newly characterized proteins are conserved in mammals, including several orthologs known to be involved in human disease. Mutations in many of these genes demonstrate statistically significant mitochondrial transmission phenotypes more subtle than could be detected by traditional genetic screens or high-throughput techniques, and 47 have not been previously localized to mitochondria. We further characterized a subset of these genes using growth profiling and dual immunofluorescence, which identified genes specifically required for aerobic respiration and an uncharacterized cytoplasmic protein required for normal mitochondrial motility. Our results demonstrate that by leveraging computational analysis to direct quantitative experimental assays, we have characterized mutants with subtle mitochondrial defects whose phenotypes were undetected by high-throughput methods.
Hess
David C
DC
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ, USA.
Myers
Chad L
CL
Huttenhower
Curtis
C
Hibbs
Matthew A
MA
Hayes
Alicia P
AP
Paw
Jadine
J
Clore
John J
JJ
Mendoza
Rosa M
RM
Luis
Bryan San
BS
Nislow
Corey
C
Giaever
Guri
G
Costanzo
Michael
M
Troyanskaya
Olga G
OG
Caudy
Amy A
AA
D019069:Cell Respiration / Q000235:genetics; D003593:Cytoplasm / Q000737:chemistry; D050259:Genes, Mitochondrial; D008928:Mitochondria / Q000235:genetics*; D024101:Mitochondrial Proteins; D050505:Mutant Proteins; D009154:Mutation; D011506:Proteins / Q000235:genetics / Q000502:physiology*; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure*
D024101:Mitochondrial Proteins; D050505:Mutant Proteins; D011506:Proteins