Dysbindin regulates the transcriptional level of myristoylated alanine-rich protein kinase C substrate via the interaction with NF-YB in mice brain.
BACKGROUND
An accumulating body of evidence suggests that Dtnbp1 (Dysbindin) is a key susceptibility gene for schizophrenia. Using the yeast-two-hybrid screening system, we examined the candidate proteins interacting with Dysbindin and revealed one of these candidates to be the transcription factor NF-YB.


METHODS
We employed an immunoprecipitation (IP) assay to demonstrate the Dysbindin-NF-YB interaction. DNA chips were used to screen for altered expression of genes in cells in which Dysbindin or NF-YB was down regulated, while Chromatin IP and Reporter assays were used to confirm the involvement of these genes in transcription of Myristoylated alanine-rich protein kinase C substrate (MARCKS). The sdy mutant mice with a deletion in Dysbindin, which exhibit behavioral abnormalities, and wild-type DBA2J mice were used to investigate MARCKS expression.


RESULTS
We revealed an interaction between Dysbindin and NF-YB. DNA chips showed that MARCKS expression was increased in both Dysbindin knockdown cells and NF-YB knockdown cells, and Chromatin IP revealed interaction of these proteins at the MARCKS promoter region. Reporter assay results suggested functional involvement of the interaction between Dysbindin and NF-YB in MARCKS transcription levels, via the CCAAT motif which is a NF-YB binding sequence. MARCKS expression was increased in sdy mutant mice when compared to wild-type mice.


CONCLUSIONS
These findings suggest that abnormal expression of MARCKS via dysfunction of Dysbindin might cause impairment of neural transmission and abnormal synaptogenesis. Our results should provide new insights into the mechanisms of neuronal development and the pathogenesis of schizophrenia.
Okuda
Hiroaki
H
Department of Anatomy and Neuroscience, Graduate School of Medicine, Osaka University, Osaka, Japan.
Kuwahara
Ryusuke
R
Matsuzaki
Shinsuke
S
Miyata
Shingo
S
Kumamoto
Natsuko
N
Hattori
Tsuyoshi
T
Shimizu
Shoko
S
Yamada
Kohei
K
Kawamoto
Keisuke
K
Hashimoto
Ryota
R
Takeda
Masatoshi
M
Katayama
Taiichi
T
Tohyama
Masaya
M
D000818:Animals; D001483:Base Sequence; D023081:CCAAT-Binding Factor / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins / Q000502:physiology*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D000074765:Dysbindin; D049029:Dystrophin-Associated Proteins; D006801:Humans; D007150:Immunohistochemistry; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D000076250:Myristoylated Alanine-Rich C Kinase Substrate; D014158:Transcription, Genetic / Q000502:physiology*
D023081:CCAAT-Binding Factor; D002352:Carrier Proteins; D017931:DNA Primers; C498992:Dtnbp1 protein, mouse; D000074765:Dysbindin; D049029:Dystrophin-Associated Proteins; D047908:Intracellular Signaling Peptides and Proteins; C000619710:MARCKS protein, human; C000619711:Marcks protein, mouse; D008565:Membrane Proteins; C492062:Nfyb protein, mouse; D000076250:Myristoylated Alanine-Rich C Kinase Substrate