Cell survival under nutrient stress is dependent on metabolic conditions regulated by Akt and not by autophagic vacuoles.
Akt activation assists tumor cell survival and promotes resistance to chemotherapy. Here we show that constitutively active Akt (CA-Akt) cells are highly sensitized to cell death induced by nutrient and growth factor deprivation, whereas dominant-negative Akt (DN-Akt) cells have a high rate of survival. The content of autophagosomes in starved CA-Akt cells was high, while DN-Akt cells expressed autophagic vacuoles constitutively, independently of nutrition conditions. Thus Akt down-regulation and downstream events can induce autophagosomes which were not directly determinants of cell death. Biochemical analysis in Akt-mutated cells show that (i) Akt and mTOR proteins were degraded more rapidly than the housekeeping proteins, (ii) mTOR phosphorylation at position Thr(2446) was relatively high in DN-Akt and low in CA-Akt cells, induced by starvation in mock cells only, which suggests reduced autoregulation of these pathways in Akt-mutated cells, (iii) both protein synthesis and protein degradation were significantly higher in starved CA-Akt cells than in starved DN-Akt cells or mock cells. In conclusion, constitutively active Akt, unable to control synthesis and wasting of proteins, accelerates the death of starved cells.
Bruno
P
P
Department of Pharmacology, University of Milan, Via Vanvitelli 32, 20129 Milan, Italy.
Calastretti
A
A
Priulla
M
M
Asnaghi
L
L
Scarlatti
F
F
Nicolin
A
A
Canti
G
G
D017209:Apoptosis; D001343:Autophagy; D002460:Cell Line; D002470:Cell Survival; D016895:Culture Media, Serum-Free; D006801:Humans; D009154:Mutation; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D058570:TOR Serine-Threonine Kinases; D014617:Vacuoles / Q000378:metabolism
D016895:Culture Media, Serum-Free; D011506:Proteins; D011494:Protein Kinases; C546842:MTOR protein, human; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases