A quantitative proteomic analysis of mitochondrial participation in p19 cell neuronal differentiation.
A quantitative proteomic analysis of changes in protein expression accompanying the differentiation of P19 mouse embryonal carcinoma cells into neuron-like cells using isobaric tag technology coupled with LC-MS/MS revealed protein changes reflecting withdrawal from the cell cycle accompanied by a dynamic reorganization of the cytoskeleton and an up-regulation of mitochondrial biogenesis. Further study of quantitative changes in abundance of individual proteins in a purified mitochondrial fraction showed that most mitochondrial proteins increased significantly in abundance. A set of chaperone proteins did not participate in this increase, suggesting that neuron-like cells are relatively deficient in mitochondrial chaperones. We developed a procedure to account for differences in recovery of mitochondrial proteins during purification of organelles from distinct cell or tissue sources. Proteomic data supported by RT-PCR analysis suggests that enhanced mitochondrial biogenesis during neuronal differentiation may reflect a large increase in expression of PGC-1alpha combined with down-regulation of its negative regulator, p160 Mybbp1a.
Watkins
Jermel
J
Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, New York 11794-8651, USA.
Basu
Siddhartha
S
Bogenhagen
Daniel F
DF
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D004268:DNA-Binding Proteins; D020869:Gene Expression Profiling; D051379:Mice; D008928:Mitochondria / Q000502:physiology*; D024101:Mitochondrial Proteins / Q000235:genetics; D009474:Neurons / Q000166:cytology*; D009687:Nuclear Proteins / Q000235:genetics; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; D011506:Proteins / Q000032:analysis / Q000235:genetics*; D040901:Proteomics / Q000379:methods*; D016601:RNA-Binding Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053719:Tandem Mass Spectrometry; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D024101:Mitochondrial Proteins; C110421:Mybbp1a protein, mouse; D009687:Nuclear Proteins; D000071248:Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha; C485290:Ppargc1a protein, mouse; D011506:Proteins; D016601:RNA-Binding Proteins; D015534:Trans-Activators; D014157:Transcription Factors