Comparative analysis of brain proteins from p53-deficient mice by two-dimensional electrophoresis.
p53 is a tumor suppressor protein that regulates many cellular processes including the cell cycle, DNA repair, and apoptosis. It also serves as a critical regulator of neuronal apoptosis in the central nervous system (CNS). To elucidate the role of p53 in the CNS, brain proteins of p53 knock-out mice (p53-/-) were analyzed by two-dimensional gel electrophoresis (2-DE) and compared with those from p53 wild type (p53+/+) mice. Six types of brain tissue (temporal cortex, cerebellum, hippocampus, striatum, olfactory bulb, and cervical spinal cord) and other control tissues (lung and blood) from 18-week-old non-stress-induced mice were analyzed. The morphology of brains from p53-/- mice appeared to be normal and identical to that of p53+/+ mice, although lungs showed diffuse tumors that may have been caused by p53 deficiency. Comparative 2-D gel analysis showed that, on average, 7 of 886 spots from brain tissue were p53-/- specific, whereas 12 of 1008 spots from lung tissue were p53-/- specific. N-terminal amino acid sequence was determined for p53-/- specific proteins. In all brain tissues from p53-/- mice, a newly identified mouse mitochondrial NADH-ubiquinone oxidoreductase 24 kDa subunit showed decreased expression, and apolipoprotein A1 acidic forms showed increased expression. In addition, brain-type creatine kinase B chain and tubulin beta-5 N-terminal fragment were increased in the p53-/- cerebellum, and a new protein in mouse, hydroxyacylglutathione hydrolase (glyoxalase II) was decreased in the temporal cortex of p53-/- mice. The alterations in protein expression identified in this study may imply a p53-related brain function. This is the first proteomic analysis on the p53-/- mouse brain, and further information based on this study will provide new insights into the p53 function in the CNS.
Araki
N
N
Department of Tumor Genetics and Biology, Kumamoto University School of Medicine, Japan. nori@gpo.kumamoto-u.ac.jp
Morimasa
T
T
Sakai
T
T
Tokuoh
H
H
Yunoue
S
S
Kamo
M
M
Miyazaki
K
K
Abe
K
K
Saya
H
H
Tsugita
A
A
D000595:Amino Acid Sequence; D000818:Animals; D016632:Apolipoprotein A-I / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D003402:Creatine Kinase / Q000378:metabolism; D042967:Electron Transport Complex I; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D013869:Thiolester Hydrolases / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D000099301:Hydroxyacylglutathione Hydrolase
D016632:Apolipoprotein A-I; D003402:Creatine Kinase; D042967:Electron Transport Complex I; D009247:NADH, NADPH Oxidoreductases; D009419:Nerve Tissue Proteins; D013869:Thiolester Hydrolases; D014404:Tubulin; D016159:Tumor Suppressor Protein p53; D000099301:Hydroxyacylglutathione Hydrolase