title: Delayed correlation of mRNA and protein expression in rapamycin-treated cells and a role for Ggc1 in cellular sensitivity to rapamycin.
abstract: To identify new molecular targets of rapamycin, an anticancer and immunosuppressive drug, we analyzed temporal changes in yeast over 6 h in response to rapamycin at the transcriptome and proteome levels and integrated the expression patterns with functional profiling. We show that the integration of transcriptomics, proteomics, and functional data sets provides novel insights into the molecular mechanisms of rapamycin action. We first observed a temporal delay in the correlation of mRNA and protein expression where mRNA expression at 1 and 2 h correlated best with protein expression changes after 6 h of rapamycin treatment. This was especially the case for the inhibition of ribosome biogenesis and induction of heat shock and autophagy essential to promote the cellular sensitivity to rapamycin. However, increased levels of vacuolar protease could enhance resistance to rapamycin. Of the 85 proteins identified as statistically significantly changing in abundance, most of the proteins that decreased in abundance were correlated with a decrease in mRNA expression. However, of the 56 proteins increasing in abundance, 26 were not correlated with an increase in mRNA expression. These protein changes were correlated with unchanged or down-regulated mRNA expression. These proteins, involved in mitochondrial genome maintenance, endocytosis, or drug export, represent new candidates effecting rapamycin action whose expression might be post-transcriptionally or post-translationally regulated. We identified GGC1, a mitochondrial GTP/GDP carrier, as a new component of the rapamycin/target of rapamycin (TOR) signaling pathway. We determined that the protein product of GGC1 was stabilized in the presence of rapamycin, and the deletion of the GGC1 enhanced growth fitness in the presence of rapamycin. A dynamic mRNA expression analysis of Deltaggc1 and wild-type cells treated with rapamycin revealed a key role for Ggc1p in the regulation of ribosome biogenesis and cell cycle progression under TOR control.
authors: Stowers Institute for Medical Research, Kansas City, Missouri 64110, USA.
authors: Marjorie L
authors: ML
authors: Fournier
authors: Ariel
authors: A
authors: Paulson
authors: Norman
authors: N
authors: Pavelka
authors: Amber L
authors: AL
authors: Mosley
authors: Karin
authors: K
authors: Gaudenz
authors: William D
authors: WD
authors: Bradford
authors: Earl
authors: E
authors: Glynn
authors: Hua
authors: H
authors: Li
authors: Mihaela E
authors: ME
authors: Sardiu
authors: Brian
authors: B
authors: Fleharty
authors: Christopher
authors: C
authors: Seidel
authors: Laurence
authors: L
authors: Florens
authors: Michael P
authors: MP
authors: Washburn
mesh_terms: D017353:Gene Deletion; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D055550:Protein Stability / Q000187:drug effects; D040901:Proteomics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D008565:Membrane Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; C105534:YHM1 protein, S cerevisiae; D020123:Sirolimus