Substitutions of potentially phosphorylatable serine residues of Bax reveal how they may regulate its interaction with mitochondria.
During apoptosis, the pro-apoptotic protein Bax relocalizes from the cytosol to the mitochondrial outer membrane. This relocalization is associated to major conformational changes, namely at the N- and C-terminal ends of the protein. Substitution of residues located at critical positions within the protein potentially stimulates or inhibits this process. In the present study, we investigated the hypothesis that phosphorylation of serine residues might trigger these conformational changes, with a focus on Ser(163) and Ser(184), which have been shown to be phosphorylatable by protein kinases GSK3beta and Akt/PKB, respectively, and on Ser(60), which is located in a consensus target sequence for PKA. Substitutions of these serine residues by alanine or aspartate were done in wild type or previously characterized Bax mutants, and the capacity of the resulting proteins to interact with mitochondria and to release cytochrome c was assayed in yeast, which provides a tool to study the function of Bax, independently of the rest of the apoptotic network. We conclude that sequential phosphorylation of these serine residues might participate in the triggering of the different conformational changes associated with Bax activation during apoptosis.
Arokium
Hubert
H
CNRS, UMR5095, Université de Bordeaux 2, 1 Rue Camille Saint-Saëns, 33077 Bordeaux, France.
Ouerfelli
Hakim
H
Velours
Gisèle
G
Camougrand
Nadine
N
Vallette
François M
FM
Manon
Stéphen
S
D000595:Amino Acid Sequence; D017209:Apoptosis; D005656:Fungal Proteins / Q000737:chemistry; D005786:Gene Expression Regulation; D038362:Glycogen Synthase Kinase 3 / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D012694:Serine / Q000737:chemistry; D051028:bcl-2-Associated X Protein / Q000235:genetics* / Q000378:metabolism*
D005656:Fungal Proteins; D051028:bcl-2-Associated X Protein; D012694:Serine; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt; D038362:Glycogen Synthase Kinase 3