A novel ninein-interaction protein, CGI-99, blocks ninein phosphorylation by GSK3beta and is highly expressed in brain tumors.
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.
Howng
Shen-Long
SL
Department of Neurosurgery, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC.
Hsu
Hui-Chun
HC
Cheng
Tai-Shan
TS
Lee
Yun-Lin
YL
Chang
Li-Kwan
LK
Lu
Pei-Jung
PJ
Hong
Yi-Ren
YR
D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D002467:Cell Nucleus / Q000378:metabolism; D003598:Cytoskeletal Proteins; D004926:Escherichia coli / Q000378:metabolism; D005333:Fetus / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014018:Tissue Distribution; D015534:Trans-Activators / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D003598:Cytoskeletal Proteins; C512806:NIN protein, human; D009687:Nuclear Proteins; C435504:RTRAF protein, human; D011994:Recombinant Proteins; D015534:Trans-Activators; D038362:Glycogen Synthase Kinase 3; D019204:GTP-Binding Proteins