title: Critical cysteines in Akt1 regulate its activity and proteasomal degradation: implications for neurodegenerative diseases.
abstract: Impaired Akt1 signaling is observed in neurodegenerative diseases, including Parkinson׳s disease (PD). In PD models oxidative modification of Akt1 leads to its dephosphorylation and consequent loss of its kinase activity. To explore the underlying mechanism we exposed Neuro2A cells to cadmium, a pan inhibitor of protein thiol disulfide oxidoreductases, including glutaredoxin 1 (Grx1), or downregulated Grx1, which led to dephosphorylation of Akt1, loss of its kinase activity, and also decreased Akt1 protein levels. Mutation of cysteines to serines at 296 and 310 in Akt1 did not affect its basal kinase activity but abolished cadmium- and Grx1 downregulation-induced reduction in Akt1 kinase activity, indicating their critical role in redox modulation of Akt1 function and turnover. Cadmium-induced decrease in phosphorylated Akt1 correlated with increased association of wild-type (WT) Akt1 with PP2A, which was absent in the C296-310S Akt1 mutant and was also abolished by N-acetylcysteine treatment. Further, increased proteasomal degradation of Akt1 by cadmium was not seen in the C296-310S Akt1 mutant, indicating that oxidation of cysteine residues facilitates degradation of WT Akt1. Moreover, preventing oxidative modification of Akt1 cysteines 296 and 310 by mutating them to serines increased the cell survival effects of Akt1. Thus, in neurodegenerative states such as PD, maintaining the thiol status of cysteines 296 and 310 in Akt1 would be critical for Akt1 kinase activity and for preventing its degradation by proteasomes. Preventing downregulation of Akt signaling not only has long-range consequences for cell survival but could also affect the multiple roles that Akt plays, including in the Akt-mTOR signaling cascade.
authors.affiliation: Centre for Neuroscience, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Faraz
authors.initials: F
authors.lastname: Ahmad
authors.affiliation: Centre for Neuroscience, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Prakash
authors.initials: P
authors.lastname: Nidadavolu
authors.affiliation: Centre for Neuroscience, Indian Institute of Science, Bangalore 560012, India.
authors.forename: Lalitha
authors.initials: L
authors.lastname: Durgadoss
authors.affiliation: Centre for Neuroscience, Indian Institute of Science, Bangalore 560012, India. Electronic address: viji@cns.iisc.ernet.in.
authors.forename: Vijayalakshmi
authors.initials: V
authors.lastname: Ravindranath
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D002147:Calmodulin / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003545:Cysteine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000235:genetics; D054477:Glutaredoxins / Q000037:antagonists & inhibitors; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D010300:Parkinson Disease / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011490:Protein Disulfide Reductase (Glutathione) / Q000037:antagonists & inhibitors; D054648:Protein Phosphatase 2 / Q000378:metabolism*; D059748:Proteolysis; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics
chemical_list: D002147:Calmodulin; C516006:Glrx protein, mouse; D054477:Glutaredoxins; D011490:Protein Disulfide Reductase (Glutathione); C514862:Akt1 protein, mouse; D051057:Proto-Oncogene Proteins c-akt; D054648:Protein Phosphatase 2; D046988:Proteasome Endopeptidase Complex; D003545:Cysteine; D000111:Acetylcysteine
keywords: Free radicals; Glutaredoxin; Oxidative stress; PP2A; Parkinson disease; Protein thiols