Document Detail


Splicing factor 3b subunit 4 binds BMPR-IA and inhibits osteochondral cell differentiation.
MedLine Citation:
PMID:  17513295     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Bone morphogenetic protein (BMP)-2/4 play critical roles in early embryogenesis and skeletal development. BMP-2/4 signals conduct into cells via two types of serine/threonine kinase receptors, known as BMPR-I (IA and IB) and BMPR-II. Here we identified splicing factor 3b subunit 4 (SF3b4) as a molecule that interacts with BMPR-IA, using a yeast two-hybrid screening with a human fetal brain cDNA library. Co-immunoprecipitation/immunoblot analysis confirmed their interaction in mammalian cells. By separation of the cell components, SF3b4 was present in the cell membrane fraction with BMPR-IA as well as in the nucleus. Overexpression of SF3b4 inhibited BMP-2-mediated osteogenic and chondrocytic differentiation of C2C12 and ATDC5 cells, respectively, and the endogenous expression level of SF3b4 decreased during differentiation in ATDC5 cells. By reporter gene assay, SF3b4 suppressed Id reporter gene activity, specific to the Smad1/5/8 pathway, but not TGFbeta-mediated reporter gene activity. Biotin labeling of the cell surface proteins followed by their immunoblot revealed that SF3b4 decreased the cell surface BMPRI-A levels. Further analysis by molecular modeling of the intracellular domain of BMPR-IA, coupled with binding studies of its several mutants, indicated that the site(s) for SF3b4 binding is not directly associated with the C-terminal lobe and the activation segment. Taken together, these results suggest that SF3b4, known to be localized in the nucleus and involved in RNA splicing, binds BMPR-IA and specifically inhibits BMP-mediated osteochondral cell differentiation.
Authors:
Hiroki Watanabe; Masafumi Shionyu; Tomoatsu Kimura; Koji Kimata; Hideto Watanabe
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-05-18
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  282     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2007 Jul 
Date Detail:
Created Date:  2007-07-09     Completed Date:  2007-09-13     Revised Date:  2012-06-25    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  20728-38     Citation Subset:  IM    
Affiliation:
Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480-1195, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Binding Sites / genetics
Bone Morphogenetic Protein Receptors, Type I / biosynthesis*,  genetics
Bone Morphogenetic Protein Receptors, Type II / genetics,  metabolism
COS Cells
Cell Differentiation / physiology*
Cell Line
Cell Membrane / genetics,  metabolism
Cell Nucleus / genetics,  metabolism
Cercopithecus aethiops
Chondrogenesis / physiology*
Humans
Models, Molecular
Mutation
Osteogenesis / physiology*
Protein Binding / genetics
Protein Structure, Tertiary / genetics
RNA Splicing / physiology
RNA-Binding Proteins / genetics,  metabolism*
Signal Transduction / physiology*
Smad Proteins / genetics,  metabolism
Transforming Growth Factor beta / genetics,  metabolism
Two-Hybrid System Techniques
Chemical
Reg. No./Substance:
0/RNA-Binding Proteins; 0/SF3B4 protein, human; 0/Smad Proteins; 0/Transforming Growth Factor beta; EC 2.7.11.30/BMPR1A protein, human; EC 2.7.11.30/BMPR2 protein, human; EC 2.7.11.30/Bone Morphogenetic Protein Receptors, Type I; EC 2.7.11.30/Bone Morphogenetic Protein Receptors, Type II

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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