Implication of Akt-dependent Prp19 alpha/14-3-3beta/Cdc5L complex formation in neuronal differentiation.
PRP19alpha and CDC5L are major components of the active spliceosome. However, their association process is still unknown. Here, we demonstrated that PRP19 alpha/14-3-3beta/CDC5L complex formation is regulated by Akt during nerve growth factor (NGF)-induced neuronal differentiation of PC12 cells. Analysis of PRP19 alpha mutants revealed that the phosphorylation of PRP19 alpha at Thr 193 by Akt was critical for its binding with 14-3-3beta to translocate into the nuclei and for PRP19 alpha/14-3-3beta/CDC5L complex formation in neuronal differentiation. Forced expression of either sense PRP19 alpha or sense 14-3-3beta RNAs promoted NGF-induced neuronal differentiation, whereas down-regulation of these mRNAs showed a suppressive effect. The nonphosphorylation mutant PRP19 alpha T193A lost its binding ability with 14-3-3beta and acted as a dominant-negative mutant in neuronal differentiation. These results imply that Akt-dependent phosphorylation of PRP19 alpha at Thr193 triggers PRP19 alpha/14-3-3beta/CDC5L complex formation in the nuclei, likely to assemble the active spliceosome against neurogenic pre-mRNAs.
Urano-Tashiro
Yumiko
Y
Department of Biological Science and Technology, Faculty of Industrial Science and Technology, Tokyo University of Science, Chiba, Japan.
Sasaki
Hironobu
H
Sugawara-Kawasaki
Mioko
M
Yamada
Takeyuki
T
Sugiyama
Akinori
A
Akiyama
Hirotada
H
Kawasaki
Yasushi
Y
Tashiro
Fumio
F
D048948:14-3-3 Proteins / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002522:Chlorocebus aethiops; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D047468:Immunoprecipitation / Q000379:methods; D020932:Nerve Growth Factor / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism*; D051058:Oncogene Protein v-akt / Q000378:metabolism*; D016716:PC12 Cells; D010766:Phosphorylation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D051381:Rats; D013752:Tetracycline / Q000494:pharmacology; D013912:Threonine / Q000378:metabolism
D048948:14-3-3 Proteins; C082279:CDC15 protein; D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; D034521:Nuclear Matrix-Associated Proteins; D011500:Protein Synthesis Inhibitors; D013912:Threonine; D020932:Nerve Growth Factor; D051058:Oncogene Protein v-akt; D019204:GTP-Binding Proteins; D013752:Tetracycline