title: Far Upstream Element-Binding Protein 1 Binds the 3' Untranslated Region of PKD2 and Suppresses Its Translation.
abstract: Autosomal dominant polycystic kidney disease pathogenesis can be recapitulated in animal models by gene mutations in or dosage alterations of polycystic kidney disease 1 (PKD1) or PKD2, demonstrating that too much and too little PKD1/PKD2 are both pathogenic. Gene dosage manipulation has become an appealing approach by which to compensate for loss or gain of gene function, but the mechanisms controlling PKD2 expression remain incompletely characterized. In this study, using cultured mammalian cells and dual-luciferase assays, we found that the 3' untranslated region (3'UTR) of PKD2 mRNA inhibits luciferase protein expression. We then identified nucleotides 691-1044, which we called 3FI, as the 3'UTR fragment necessary for repressing the expression of luciferase or PKD2 in this system. Using a pull-down assay and mass spectrometry we identified far upstream element-binding protein 1 (FUBP1) as a 3FI-binding protein. In vitro overexpression of FUBP1 inhibited the expression of PKD2 protein but not mRNA. In embryonic zebrafish, FUBP1 knockdown (KD) by morpholino injection increased PKD2 expression and alleviated fish tail curling caused by morpholino-mediated KD of PKD2. Conversely, FUBP1 overexpression by mRNA injection significantly increased pronephric cyst occurrence and tail curling in zebrafish embryos. Furthermore, FUBP1 binds directly to eukaryotic translation initiation factor 4E-binding protein 1, indicating a link to the translation initiation complex. These results show that FUBP1 binds 3FI in the PKD2 3'UTR to inhibit PKD2 translation, regulating zebrafish disease phenotypes associated with PKD2 KD.
authors.affiliation: Membrane Protein Disease and Cancer Research Centre, Provincial Innovation Center, Hubei University of Technology, Wuhan, China; Membrane Protein Disease Research Group, Department of Physiology.
authors.forename: Wang
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Membrane Protein Disease Research Group, Department of Physiology, Medical Research Center, Zhongnan Hospital, Wuhan University, Wuhan, China; and.
authors.forename: Fan
authors.initials: F
authors.lastname: Shen
authors.affiliation: School of Life Sciences and Technology, Tongji University, Shanghai, China.
authors.forename: Ruikun
authors.initials: R
authors.lastname: Hu
authors.affiliation: Department of Biological Sciences, and.
authors.forename: Birbickram
authors.initials: B
authors.lastname: Roy
authors.affiliation: Membrane Protein Disease Research Group, Department of Physiology.
authors.forename: JungWoo
authors.initials: J
authors.lastname: Yang
authors.affiliation: Membrane Protein Disease Research Group, Department of Physiology.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: School of Life Sciences and Technology, Tongji University, Shanghai, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Membrane Protein Disease Research Group, Department of Physiology.
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: King
authors.affiliation: Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta, Canada;
authors.forename: Consolato
authors.initials: C
authors.lastname: Sergi
authors.affiliation: Medical Research Center, Zhongnan Hospital, Wuhan University, Wuhan, China; and.
authors.forename: Song-Mei
authors.initials: SM
authors.lastname: Liu
authors.affiliation: Membrane Protein Disease Research Group, Department of Physiology.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Cordat
authors.affiliation: Membrane Protein Disease and Cancer Research Centre, Provincial Innovation Center, Hubei University of Technology, Wuhan, China;
authors.forename: Jingfeng
authors.initials: J
authors.lastname: Tang
authors.affiliation: School of Life Sciences and Technology, Tongji University, Shanghai, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Biological Sciences, and.
authors.forename: Declan
authors.initials: D
authors.lastname: Ali
authors.affiliation: Membrane Protein Disease and Cancer Research Centre, Provincial Innovation Center, Hubei University of Technology, Wuhan, China; Membrane Protein Disease Research Group, Department of Physiology, xzchen@ualberta.ca.
authors.forename: Xing-Zhen
authors.initials: XZ
authors.lastname: Chen
mesh_terms: D020413:3' Untranslated Regions / Q000502:physiology*; D000818:Animals; D002478:Cells, Cultured; D004265:DNA Helicases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins; D050396:TRPP Cation Channels / Q000235:genetics*; D015027:Zebrafish
chemical_list: D020413:3' Untranslated Regions; D004265:DNA Helicases; D004268:DNA-Binding Proteins; D016601:RNA-Binding Proteins; D050396:TRPP Cation Channels; C492258:FUBP1 protein, human
keywords: 3'UTR; ADPKD; RNA-binding protein; translational regulation; zebrafish
references.citation: Nat Med. 2008 Aug;14(8):863-8
references.pmid: 18552856
references.citation: Hum Mutat. 2014 Dec;35(12):1393-406
references.pmid: 25263802
references.citation: Nat Rev Genet. 2012 Jun;13(6):383-94
references.pmid: 22568971
references.citation: Cell. 2001 Jan 26;104(2):281-90
references.pmid: 11207368
references.citation: Hum Mol Genet. 1999 Mar;8(3):509-13
references.pmid: 9949210
references.citation: Genes Dev. 1994 Feb 15;8(4):465-80
references.pmid: 8125259
references.citation: Oncogene. 2013 Jun 13;32(24):2907-16
references.pmid: 22926519
references.citation: Am J Physiol. 1999 Jul;277(1 Pt 2):F17-25
references.pmid: 10409293
references.citation: Development. 2003 Apr;130(8):1549-64
references.pmid: 12620981
references.citation: EMBO J. 2011 Mar 2;30(5):846-58
references.pmid: 21285945
references.citation: Oncogene. 1995 Jun 1;10(11):2229-38
references.pmid: 7784068
references.citation: Trends Cardiovasc Med. 2006 Nov;16(8):292-8
references.pmid: 17055386
references.citation: Mol Cell Biol. 2005 Sep;25(18):8285-98
references.pmid: 16135816
references.citation: Dev Dyn. 2010 Feb;239(2):482-95
references.pmid: 20034105
references.citation: Nat Rev Mol Cell Biol. 2010 Feb;11(2):113-27
references.pmid: 20094052
references.citation: J Biol Chem. 2004 Jul 9;279(28):29728-39
references.pmid: 15123714
references.citation: Development. 1998 Dec;125(23):4655-67
references.pmid: 9806915
references.citation: Am J Pathol. 2006 Jan;168(1):205-20
references.pmid: 16400024
references.citation: Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6965-70
references.pmid: 9192675
references.citation: Development. 2010 Apr;137(7):1107-16
references.pmid: 20215348
references.citation: J Biol Chem. 2008 Nov 14;283(46):31559-66
references.pmid: 18782757
references.citation: Oncogene. 2009 Mar 26;28(12):1529-36
references.pmid: 19219071
references.citation: BMC Cancer. 2008 Dec 16;8:369
references.pmid: 19087307
references.citation: J Biol Chem. 2004 Mar 26;279(13):12974-9
references.pmid: 14711832
references.citation: Anal Biochem. 2005 Nov 15;346(2):210-6
references.pmid: 16212928
references.citation: Nat Genet. 2003 Jul;34(3):330-6
references.pmid: 12819782
references.citation: Cell Signal. 2004 Oct;16(10):1113-21
references.pmid: 15240006
references.citation: J Biol Chem. 2008 Oct 17;283(42):28471-9
references.pmid: 18701462
references.citation: FASEB J. 2013 Dec;27(12):4998-5009
references.pmid: 23985800
references.citation: J Am Soc Nephrol. 2011 May;22(5):791-3
references.pmid: 21474564
references.citation: Nucleic Acids Res. 1997 Mar 15;25(6):1281-8
references.pmid: 9092640
references.citation: Gene. 1998 Aug 17;216(1):1-11
references.pmid: 9714706
references.citation: Mol Cell Biol. 2000 Feb;20(3):760-9
references.pmid: 10629032
references.citation: Proteomics. 2009 Mar;9(5):1230-40
references.pmid: 19253281
references.citation: Proc Natl Acad Sci U S A. 2001 Apr 10;98(8):4288-9
references.pmid: 11296278
references.citation: Cell. 1998 Apr 17;93(2):177-88
references.pmid: 9568711
references.citation: Mol Cell Biol. 2001 Oct;21(19):6440-9
references.pmid: 11533233
references.citation: Pflugers Arch. 2005 Oct;451(1):277-85
references.pmid: 15971078
references.citation: J Biol Chem. 2005 Dec 16;280(50):41298-306
references.pmid: 16223735
references.citation: J Am Soc Nephrol. 1999 Jul;10(7):1524-9
references.pmid: 10405208
references.citation: J Clin Invest. 1997 Jan 15;99(2):194-9
references.pmid: 9005987
references.citation: J Biol Chem. 2009 Mar 13;284(11):7214-22
references.pmid: 19098310
references.citation: Kidney Int. 2002 Jan;61(1 Suppl):S125-31
references.pmid: 11841626
references.citation: Oncogene. 1996 Sep 19;13(6):1153-60
references.pmid: 8808689
references.citation: Dev Neurobiol. 2015 Feb;75(2):145-62
references.pmid: 25047640
references.citation: Comp Med. 2010 Apr;60(2):96-106
references.pmid: 20412683
references.citation: Proc Natl Acad Sci U S A. 1999 Mar 30;96(7):3934-9
references.pmid: 10097141
references.citation: Mol Biol Rep. 2010 Jul;37(6):2951-8
references.pmid: 19821056
references.citation: Proc Natl Acad Sci U S A. 2001 Jan 30;98(3):1182-7
references.pmid: 11252306
references.citation: Development. 2004 Aug;131(16):4085-93
references.pmid: 15269167
references.citation: Hum Mol Genet. 2005 Jun 15;14(12):1587-603
references.pmid: 15843396
references.citation: Annu Rev Biochem. 1999;68:913-63
references.pmid: 10872469
references.citation: Nat Cell Biol. 2005 Dec;7(12):1202-12
references.pmid: 16311606
references.citation: Eur J Hum Genet. 2005 May;13(5):649-59
references.pmid: 15770226
references.citation: Hepatology. 2009 Oct;50(4):1121-9
references.pmid: 19637194
references.citation: Nat Genet. 1997 Oct;17(2):179-81
references.pmid: 9326937
references.citation: EMBO J. 2010 Apr 7;29(7):1176-91
references.pmid: 20168298
references.citation: Hum Mol Genet. 2004 Dec 15;13(24):3069-77
references.pmid: 15496422
references.citation: Neurosci Lett. 2013 Feb 28;539:54-9
references.pmid: 23384568
references.citation: Oncogene. 2011 Jan 6;30(1):77-86
references.pmid: 20802533
references.citation: Hum Mol Genet. 2010 Apr 1;19(7):1174-89
references.pmid: 20053665
references.citation: Cell. 2007 Jun 29;129(7):1311-23
references.pmid: 17604720
references.citation: Nat Genet. 1992 Jul;1(4):235-7
references.pmid: 1338768
references.citation: Cell. 2005 Oct 7;123(1):49-63
references.pmid: 16213212