Involvement of the mouse Prp19 gene in neuronal/astroglial cell fate decisions.
The molecular mechanisms involved in neuronal/astroglial cell fate decisions during the development of the mammalian central nervous system are poorly understood. Here, we report that PRP19beta, a splice variant of mouse PRP19alpha corresponding to the yeast PRP19 protein, can function as a neuron-astroglial switch during the retinoic acid-primed neural differentiation of P19 cells. The beta-variant possesses an additional 19 amino acid residues in-frame in the N-terminal region of the alpha-variant. The forced expression of the alpha-variant RNA caused the down-regulation of oct-3/4 and nanog mRNA expression during the 12-48 h of the late-early stages of neural differentiation and was sufficient to convert P19 cells into neurons (but not glial cells) when the cells were cultured in aggregated form without retinoic acid. In contrast, the forced expression of the beta-variant RNA suppressed neuronal differentiation and conversely stimulated astroglial cell differentiation in retinoic acid-primed P19 cells. Based on yeast two-hybrid screening, cyclophilin A was identified as a specific binding partner of the beta-variant. Luciferase reporter assay mediated by the oct-3/4 promoter revealed that cyclophilin A could act as a transcriptional activator and that its activity was suppressed by the beta-variant, suggesting that cyclophilin A takes part in the induction of oct-3/4 gene expression, which might lead to neuroectodermal otx2 expression within 12 h of the immediate-early stages of retinoic acid-primed neural differentiation. These results show that the alpha-variant gene plays a pivotal role in neural differentiation and that the beta-variant participates in neuronal/astroglial cell fate decisions.
Urano
Yumiko
Y
Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Yamazaki, Noda-shi, Chiba 270-8510, Japan.
Iiduka
Masayuki
M
Sugiyama
Akinori
A
Akiyama
Hirotada
H
Uzawa
Kouji
K
Matsumoto
Gaku
G
Kawasaki
Yasushi
Y
Tashiro
Fumio
F
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D002352:Carrier Proteins / Q000502:physiology*; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D021984:Cyclophilin A / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D045643:DNA Repair Enzymes; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008954:Models, Biological; D008969:Molecular Sequence Data; D009457:Neuroglia / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D009687:Nuclear Proteins; D009841:Oligonucleotides / Q000737:chemistry; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry / Q000378:metabolism; D000072260:RNA Splicing Factors; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D017381:Spliceosomes / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D020798:Two-Hybrid System Techniques
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D009687:Nuclear Proteins; D009841:Oligonucleotides; D000072260:RNA Splicing Factors; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D012313:RNA; D008156:Luciferases; D021984:Cyclophilin A; D045643:DNA Repair Enzymes; C417041:PRPF19 protein, human