PTEN/Akt signaling controls mitochondrial respiratory capacity through 4E-BP1.
Akt, a serine/threonine kinase has been shown to stimulate glycolysis in cancer cells but its role in mitochondrial respiration is unknown. Using PTEN-knockout mouse embryonic fibroblasts (MEF(PTEN-/-)) with hyper-activated Akt as a cell model, we observed a higher respiratory capacity in MEF(PTEN-/-) compared to the wildtype (MEF(WT)). The respiratory phenotype observed in MEF(PTEN-/-) was reproduced in MEF(WT) by gene silencing of PTEN which substantiated its role in regulating mitochondrial function. The increased activities of the respiratory complexes (RCs) I, III and IV were retained in the same relative proportions as those present in MEF(WT), alluding to a possible co-ordinated regulation by PTEN/Akt. Using LY294002 (a PI3K inhibitor) and Akt inhibitor IV, we showed that the regulation of enzyme activities and protein expressions of the RCs was dependent on PI3K/Akt. There was insignificant difference in the protein expressions of mitochondrial transcription factor: peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and its downstream targets, the nuclear respiratory factor 1 (NRF-1) and mitochondrial transcription factor A (mtTFA) between MEF(PTEN-/-) and MEF(WT). Similarly, mRNA levels of the same subunits of the RCs detected in Western blots were not significantly different between MEF(PTEN-/-) and MEF(WT) suggesting that the regulation by Akt on mitochondrial function was probably not via gene transcription. On the other hand, a decrease of total 4E-BP1 with a higher expression of its phosphorylated form relative to total 4E-BP1 was found in MEF(PTEN-/-), which inferred that the regulation of mitochondrial respiratory activities by Akt was in part through this protein translation pathway. Notably, gene silencing of 4E-BP1 up-regulated the protein expressions of all RCs and the action of 4E-BP1 appeared to be specific to these mitochondrial proteins. In conclusion, PTEN inactivation bestowed a bioenergetic advantage to the cells by up-regulating mitochondrial respiratory capacity through the 4E-BP1-mediated protein translation pathway.
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, National University Health Systems, Singapore, Singapore.
Chong Kiat
CK
Goo
Hwee Ying
HY
Lim
Qin Shi
QS
Ho
Heng-Phon
HP
Too
Marie-Veronique
MV
Clement
Kim Ping
KP
Wong
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D002867:Chromones / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D020868:Gene Silencing; D008564:Membrane Potentials; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism*; D009025:Morpholines / Q000494:pharmacology; D051059:PTEN Phosphohydrolase / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D014233:Tricarboxylic Acids / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D018797:Cell Cycle Proteins; D002867:Chromones; C087000:EIF4EBP1 protein, human; D004791:Enzyme Inhibitors; D009025:Morpholines; D010750:Phosphoproteins; D014233:Tricarboxylic Acids; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D000255:Adenosine Triphosphate; D003576:Electron Transport Complex IV; D051057:Proto-Oncogene Proteins c-akt; D051059:PTEN Phosphohydrolase; C487757:Pten protein, mouse