Abstract
To explore more hNinein interacting proteins, the yeast two-hybrid screening using ninein C-terminal domain as bait protein was performed. One novel gene, CGI-99, was demonstrated to associate with hNinein in the yeast two-hybrid method and in vitro GST pull-down assay. Molecular characterization also showed that CGI-99 possessed a transcriptional activity at the N-terminal. In addition, CGI-99 formed a dimer with the C-terminal, which overlapped with hNinein binding site. In kinase assay, CGI-99 binds to hNinein and completely blocks the phosphorylation of hNinein by GSK3beta. Moreover, CGI-99 was highly expressed in all brain tumors which is in agreement with the Northern blot analysis. Taken together, we have isolated a novel protein CGI-99, which may be involved in the functional regulation of human ninein in the centrosome structure and may also be important in brain development and tumorigenesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adult
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Amino Acid Sequence
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Base Sequence
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Blotting, Western
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Brain Neoplasms / metabolism*
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Brain Neoplasms / pathology
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Cell Nucleus / metabolism
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Cytoskeletal Proteins
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Escherichia coli / metabolism
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Fetus / metabolism
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GTP-Binding Proteins / chemistry
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GTP-Binding Proteins / genetics
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GTP-Binding Proteins / metabolism*
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Gene Expression
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Glycogen Synthase Kinase 3 / antagonists & inhibitors
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Glycogen Synthase Kinase 3 / metabolism*
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HeLa Cells
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Humans
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Male
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Molecular Sequence Data
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Nuclear Proteins
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Phosphorylation
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Tissue Distribution
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Trans-Activators / biosynthesis
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Trans-Activators / chemistry
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Trans-Activators / genetics
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Trans-Activators / metabolism*
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Transfection
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Two-Hybrid System Techniques
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Yeasts / genetics
Substances
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Cytoskeletal Proteins
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NIN protein, human
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Nuclear Proteins
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RTRAF protein, human
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Recombinant Proteins
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Trans-Activators
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Glycogen Synthase Kinase 3
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GTP-Binding Proteins