title: HPat a decapping activator interacting with the miRNA effector complex.
abstract: Animal miRNAs commonly mediate mRNA degradation and/or translational repression by binding to their target mRNAs. Key factors for miRNA-mediated mRNA degradation are the components of the miRNA effector complex (AGO1 and GW182) and the general mRNA degradation machinery (deadenylation and decapping enzymes). The CCR4-NOT1 complex required for the deadenylation of target mRNAs is directly recruited to the miRNA effector complex. However, it is unclear whether the following decapping step is only a consequence of deadenylation occurring independent of the miRNA effector complex or e.g. decapping activators can get recruited to the miRNA effector complex. In this study we performed split-affinity purifications in Drosophila cells and provide evidence for the interaction of the decapping activator HPat with the miRNA effector complex. Furthermore, in knockdown analysis of various mRNA degradation factors we demonstrate the importance of NOT1 for this interaction. This suggests that deadenylation and/or the recruitment of NOT1 protein precedes the association of HPat with the miRNA effector complex. Since HPat couples deadenylation and decapping, the recruitment of HPat to the miRNA effector complex provides a mechanism to commit the mRNA target for degradation.
authors: Max F. Perutz Laboratories, University of Vienna, Department of Microbiology, Immunbiology and Genetics, Vienna, Austria.
authors: Elisabeth
authors: E
authors: Barišić-Jäger
authors: Izabela
authors: I
authors: Kręcioch
authors: Stefanie
authors: S
authors: Hosiner
authors: Sanja
authors: S
authors: Antic
authors: Silke
authors: S
authors: Dorner
mesh_terms: D000818:Animals; D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism; D020169:Caspases; D002460:Cell Line; D002846:Chromatography, Affinity; D029721:Drosophila Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D047468:Immunoprecipitation; D035683:MicroRNAs / Q000502:physiology*; D025941:Protein Interaction Mapping; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000302:isolation & purification / Q000502:physiology*
chemical_list: C489654:AGO1 protein, Drosophila; D060565:Argonaute Proteins; D029721:Drosophila Proteins; C513405:Gw protein, Drosophila; D035683:MicroRNAs; C549399:Patr-1 protein, Drosophila; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D005095:Exoribonucleases; C487208:pcm protein, Drosophila; D020169:Caspases; C103568:Dcp-1 protein, Drosophila
references: Mol Cell. 2010 Sep 10;39(5):773-83
references: 20832728
references: Cell. 2005 Sep 23;122(6):875-86
references: 16179257
references: Nat Struct Mol Biol. 2008 Apr;15(4):346-53
references: 18345015
references: Proc Natl Acad Sci U S A. 2006 Aug 8;103(32):11880-5
references: 16882716
references: J Biol Chem. 2010 Apr 30;285(18):13599-606
references: 20181956
references: Nat Struct Mol Biol. 2012 Jun 05;19(6):586-93
references: 22664986
references: J Immunol Methods. 2007 Jan 10;318(1-2):11-9
references: 17141265
references: Curr Biol. 2005 Dec 6;15(23):2149-55
references: 16289642
references: J Cell Biol. 2010 Apr 19;189(2):289-302
references: 20404111
references: Mol Cell Biol. 2008 Feb;28(4):1298-312
references: 18086885
references: Biochim Biophys Acta. 2007 Dec;1773(12):1786-92
references: 17936923
references: Annu Rev Biochem. 2004;73:861-90
references: 15189161
references: Nat Rev Genet. 2011 Feb;12(2):99-110
references: 21245828
references: RNA Biol. 2010 May-Jun;7(3):381-5
references: 20458171
references: Nat Struct Mol Biol. 2011 Oct 07;18(11):1211-7
references: 21984185
references: Nat Struct Mol Biol. 2009 Nov;16(11):1160-6
references: 19838187
references: J Cell Sci. 2012 Dec 15;125(Pt 24):6105-16
references: 23097047
references: Mol Cell Biol. 2010 Mar;30(6):1486-94
references: 20065043
references: Nat Rev Mol Cell Biol. 2007 Feb;8(2):113-26
references: 17245413
references: Mol Cell. 2009 Sep 24;35(6):868-80
references: 19716330
references: EMBO J. 2004 Jul 21;23(14):2862-71
references: 15215893
references: RNA. 2008 Dec;14(12):2580-96
references: 18978028
references: Mol Cell Biol. 2009 Dec;29(23):6220-31
references: 19797087
references: Nat Struct Mol Biol. 2010 Feb;17(2):238-40
references: 20098421
references: Mol Cell. 2012 Dec 28;48(6):825-36
references: 23123195
references: Nucleic Acids Res. 2013 Jan;41(2):978-94
references: 23172285
references: Mol Cell. 2011 Oct 7;44(1):120-33
references: 21981923
references: Nucleic Acids Res. 2012 Jul;40(12):5651-65
references: 22402495
references: RNA. 2010 Jul;16(7):1356-70
references: 20504953
references: Nat Struct Mol Biol. 2005 Dec;12(12):1054-63
references: 16284618
references: Nat Struct Mol Biol. 2011 Oct 07;18(11):1218-26
references: 21984184
references: Science. 2006 Apr 7;312(5770):75-9
references: 16484454
references: Methods. 2009 Jul;48(3):306-10
references: 19398006
references: Mol Cell Biol. 2010 Sep;30(17):4308-23
references: 20584987
references: EMBO Rep. 2007 Nov;8(11):1052-60
references: 17932509
references: Methods. 2001 Dec;25(4):402-8
references: 11846609
references: Genes Dev. 2006 Jul 15;20(14):1885-98
references: 16815998
references: Mol Cell. 2009 Apr 10;34(1):58-67
references: 19268617
references: Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4034-9
references: 16495412
references: Genes Dev. 2007 Oct 15;21(20):2558-70
references: 17901217
references: EMBO J. 2000 Apr 3;19(7):1661-71
references: 10747033
references: Nature. 2000 Mar 30;404(6777):515-8
references: 10761922
references: Mol Cell Biol. 2000 Aug;20(16):5939-46
references: 10913177
references: Nat Struct Mol Biol. 2012 Dec;19(12):1324-31
references: 23142987
references: Mol Cell. 2007 Nov 30;28(4):598-613
references: 18042455
references: RNA. 2009 Jan;15(1):21-32
references: 19029310