Two PABPC1-binding Sites in GW182 Proteins Promote MiRNA-mediated Gene Silencing

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2010 Nov 11

PMID 21063388

Citations 61

Authors

Eric Huntzinger

Joerg E Braun

Susanne Heimstadt

Latifa Zekri

Elisa Izaurralde

Affiliations

Soon will be listed here.

Abstract

miRNA-mediated gene silencing requires the GW182 proteins, which are characterized by an N-terminal domain that interacts with Argonaute proteins (AGOs), and a C-terminal silencing domain (SD). In Drosophila melanogaster (Dm) GW182 and a human (Hs) orthologue, TNRC6C, the SD was previously shown to interact with the cytoplasmic poly(A)-binding protein (PABPC1). Here, we show that two regions of GW182 proteins interact with PABPC1: the first contains a PABP-interacting motif 2 (PAM2; as shown before for TNRC6C) and the second contains the M2 and C-terminal sequences in the SD. The latter mediates indirect binding to the PABPC1 N-terminal domain. In D. melanogaster cells, the second binding site dominates; however, in HsTNRC6A-C the PAM2 motif is essential for binding to both Hs and DmPABPC1. Accordingly, a single amino acid substitution in the TNRC6A-C PAM2 motif abolishes the interaction with PABPC1. This mutation also impairs TNRC6s silencing activity. Our findings reveal that despite species-specific differences in the relative strength of the PABPC1-binding sites, the interaction between GW182 proteins and PABPC1 is critical for miRNA-mediated silencing in animal cells.

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References

Kozlov G, De Crescenzo G, Lim N, Siddiqui N, Fantus D, Kahvejian A . Structural basis of ligand recognition by PABC, a highly specific peptide-binding domain found in poly(A)-binding protein and a HECT ubiquitin ligase. EMBO J. 2003; 23(2):272-81. PMC: 1271756. DOI: 10.1038/sj.emboj.7600048. View

Eulalio A, Huntzinger E, Izaurralde E . GW182 interaction with Argonaute is essential for miRNA-mediated translational repression and mRNA decay. Nat Struct Mol Biol. 2008; 15(4):346-53. DOI: 10.1038/nsmb.1405. View

Kozlov G, Safaee N, Rosenauer A, Gehring K . Structural basis of binding of P-body-associated proteins GW182 and ataxin-2 by the Mlle domain of poly(A)-binding protein. J Biol Chem. 2010; 285(18):13599-606. PMC: 2859521. DOI: 10.1074/jbc.M109.089540. View

Albrecht M, Lengauer T . Survey on the PABC recognition motif PAM2. Biochem Biophys Res Commun. 2004; 316(1):129-38. DOI: 10.1016/j.bbrc.2004.02.024. View

Ding L, Han M . GW182 family proteins are crucial for microRNA-mediated gene silencing. Trends Cell Biol. 2007; 17(8):411-6. DOI: 10.1016/j.tcb.2007.06.003. View

Karim M, Svitkin Y, Kahvejian A, De Crescenzo G, Costa-Mattioli M, Sonenberg N . A mechanism of translational repression by competition of Paip2 with eIF4G for poly(A) binding protein (PABP) binding. Proc Natl Acad Sci U S A. 2006; 103(25):9494-9. PMC: 1480435. DOI: 10.1073/pnas.0603701103. View

Eulalio A, Helms S, Fritzsch C, Fauser M, Izaurralde E . A C-terminal silencing domain in GW182 is essential for miRNA function. RNA. 2009; 15(6):1067-77. PMC: 2685512. DOI: 10.1261/rna.1605509. View

Ezzeddine N, Chang T, Zhu W, Yamashita A, Chen C, Zhong Z . Human TOB, an antiproliferative transcription factor, is a poly(A)-binding protein-dependent positive regulator of cytoplasmic mRNA deadenylation. Mol Cell Biol. 2007; 27(22):7791-801. PMC: 2169145. DOI: 10.1128/MCB.01254-07. View

Till S, Lejeune E, Thermann R, Bortfeld M, Hothorn M, Enderle D . A conserved motif in Argonaute-interacting proteins mediates functional interactions through the Argonaute PIWI domain. Nat Struct Mol Biol. 2007; 14(10):897-903. DOI: 10.1038/nsmb1302. View

10.

Roy G, De Crescenzo G, Khaleghpour K, Kahvejian A, OConnor-Mccourt M, Sonenberg N . Paip1 interacts with poly(A) binding protein through two independent binding motifs. Mol Cell Biol. 2002; 22(11):3769-82. PMC: 133836. DOI: 10.1128/MCB.22.11.3769-3782.2002. View

11.

Walters R, Bradrick S, Gromeier M . Poly(A)-binding protein modulates mRNA susceptibility to cap-dependent miRNA-mediated repression. RNA. 2009; 16(1):239-50. PMC: 2802033. DOI: 10.1261/rna.1795410. View

12.

Behm-Ansmant I, Rehwinkel J, Doerks T, Stark A, Bork P, Izaurralde E . mRNA degradation by miRNAs and GW182 requires both CCR4:NOT deadenylase and DCP1:DCP2 decapping complexes. Genes Dev. 2006; 20(14):1885-98. PMC: 1522082. DOI: 10.1101/gad.1424106. View

13.

Eulalio A, Tritschler F, Izaurralde E . The GW182 protein family in animal cells: new insights into domains required for miRNA-mediated gene silencing. RNA. 2009; 15(8):1433-42. PMC: 2714752. DOI: 10.1261/rna.1703809. View

14.

Derry M, Yanagiya A, Martineau Y, Sonenberg N . Regulation of poly(A)-binding protein through PABP-interacting proteins. Cold Spring Harb Symp Quant Biol. 2007; 71:537-43. DOI: 10.1101/sqb.2006.71.061. View

15.

Fabian M, Mathonnet G, Sundermeier T, Mathys H, Zipprich J, Svitkin Y . Mammalian miRNA RISC recruits CAF1 and PABP to affect PABP-dependent deadenylation. Mol Cell. 2009; 35(6):868-80. PMC: 2803087. DOI: 10.1016/j.molcel.2009.08.004. View

16.

Jinek M, Fabian M, Coyle S, Sonenberg N, Doudna J . Structural insights into the human GW182-PABC interaction in microRNA-mediated deadenylation. Nat Struct Mol Biol. 2010; 17(2):238-40. PMC: 2920127. DOI: 10.1038/nsmb.1768. View

17.

Zekri L, Huntzinger E, Heimstadt S, Izaurralde E . The silencing domain of GW182 interacts with PABPC1 to promote translational repression and degradation of microRNA targets and is required for target release. Mol Cell Biol. 2009; 29(23):6220-31. PMC: 2786699. DOI: 10.1128/MCB.01081-09. View

18.

Lazzaretti D, Tournier I, Izaurralde E . The C-terminal domains of human TNRC6A, TNRC6B, and TNRC6C silence bound transcripts independently of Argonaute proteins. RNA. 2009; 15(6):1059-66. PMC: 2685519. DOI: 10.1261/rna.1606309. View

19.

Behm-Ansmant I, Gatfield D, Rehwinkel J, Hilgers V, Izaurralde E . A conserved role for cytoplasmic poly(A)-binding protein 1 (PABPC1) in nonsense-mediated mRNA decay. EMBO J. 2007; 26(6):1591-601. PMC: 1829367. DOI: 10.1038/sj.emboj.7601588. View

20.

Khaleghpour K, Kahvejian A, de Crescenzo G, Roy G, Svitkin Y, Imataka H . Dual interactions of the translational repressor Paip2 with poly(A) binding protein. Mol Cell Biol. 2001; 21(15):5200-13. PMC: 87244. DOI: 10.1128/MCB.21.15.5200-5213.2001. View