FUBP3 interacts with FGF9 3' microsatellite and positively regulates FGF9 translation.
A TG microsatellite in the 3'-untranslated region (UTR) of FGF9 mRNA has previously been shown to modulate FGF9 expression. In the present study, we investigate the possible interacting protein that binds to FGF9 3'-UTR UG-repeat and study the mechanism underlying this protein-RNA interaction. We first applied RNA pull-down assays and LC-MS analysis to identify proteins associated with this repetitive sequence. Among the identified proteins, FUBP3 specifically bound to the synthetic (UG)(15) oligoribonucleotide as shown by supershift in RNA-EMSA experiments. The endogenous FGF9 protein was upregulated in response to transient overexpression and downregulated after knockdown of FUBP3 in HEK293 cells. As the relative levels of FGF9 mRNA were similar in these two conditions, and the depletion of FUBP3 had no effect on the turn-over rate of FGF9 mRNA, these data suggested that FUBP3 regulates FGF9 expression at the post-transcriptional level. Further examination using ribosome complex pull-down assay showed overexpression of FUBP3 promotes FGF9 expression. In contrast, polyribosome-associated FGF9 mRNA decreased significantly in FUBP3-knockdown HEK293 cells. Finally, reporter assay suggested a synergistic effect of the (UG)-motif with FUBP3 to fine-tune the expression of FGF9. Altogether, results from this study showed the novel RNA-binding property of FUBP3 and the interaction between FUBP3 and FGF9 3'-UTR UG-repeat promoting FGF9 mRNA translation.
Institute of Molecular Medicine, National Cheng Kung University Medical College, Tainan, Taiwan, ROC.
Bing-Huang
BH
Gau
Tsung-Ming
TM
Chen
Yu-Heng J
YH
Shih
H Sunny
HS
Sun
D020413:3' Untranslated Regions; D001483:Base Sequence; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D051525:Fibroblast Growth Factor 9 / Q000096:biosynthesis / Q000235:genetics*; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D009843:Oligoribonucleotides / Q000378:metabolism; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*
D020413:3' Untranslated Regions; D004268:DNA-Binding Proteins; C496459:FGF9 protein, human; C492259:FUBP3 protein, human; D051525:Fibroblast Growth Factor 9; D009843:Oligoribonucleotides; D016601:RNA-Binding Proteins; D014157:Transcription Factors
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