FMIP controls the adipocyte lineage commitment of C2C12 cells by downmodulation of C/EBP alpha.
Fms interacting protein (FMIP) is a substrate for Fms tyrosine kinase, and a nuclear/cytoplasm shuttling protein with a leucine zipper. As the phosphorylation of FMIP is observed in insulin-stimulated preadipocytes, we examined the role of FMIP in adipocyte differentiation, using the mesenchymal multipotent stem cells, C2C12 cells, that can differentiate into adipocytes, muscle cells and osteoblasts. Ectopic expression of FMIP in C2C12 impairs the adipocyte differentiation induced by treatment with insulin, dexamethasone and 3-isobutyl-1-methylxanthine. These cells exhibit muscle phenotype with multinuclear morphology. Furthermore, knockdown of endogenous FMIP expression by small interfering RNA improves adipocytic lineage commitment of C2C12 cells, while impairing muscle differentiation. Upon stimulation with insulin, CCAAT/enhancer binding protein (C/EBP)beta, but not C/EBPalpha, is upregulated in cells expressing ectopic FMIP, whereas in FMIP knockdown cells, C/EBPalpha is constitutively expressed. Ectopic expression of C/EBPalpha counteracts the effects of FMIP, whereas C/EBPalpha knockdown partially mimics the effects of FMIP in this system. Northern blot analysis and reverse transcriptase-polymerase chain reaction study reveal that ectopic FMIP-expressing cells do not contain the polyadenylated C/EBPalpha mRNA, but contain the C/EBPalpha pre-mRNA, suggesting that FMIP plays a role in RNA processing and/or export. Indeed, a member of the THO complex that plays a role in mRNA export, THOC1, is co-precipitated with FMIP. The data we have acquired on FMIP suggest that it is a target for tyrosine kinase receptors that potentiate mRNA export.
Mancini
A
A
Institut fuer Biochemie, Medizinische Hochschule Hannover, Hannover, Germany.
El Bounkari
O
O
Norrenbrock
A-F
AF
Scherr
M
M
Schaefer
D
D
Eder
M
M
Banham
A H
AH
Pulford
K
K
Lyne
L
L
Whetton
A D
AD
Tamura
T
T
D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage / Q000502:physiology*; D015536:Down-Regulation / Q000502:physiology*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D051379:Mice; D032342:Muscle Cells / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D012322:RNA Precursors / Q000096:biosynthesis; D034741:RNA, Small Interfering / Q000502:physiology
D022763:CCAAT-Enhancer-Binding Protein-alpha; C402364:Fmip protein, mouse; D047908:Intracellular Signaling Peptides and Proteins; D012322:RNA Precursors; D034741:RNA, Small Interfering