A Divergent Sm Fold in EDC3 Proteins Mediates DCP1 Binding and P-Body Targeting

Tritschler, Felix; Eulálio, Ana; Truffault, Vincent; Hartmann, M.D.; Helms, Sigrun; Schmidt, Steffen; Coles, Murray; Izaurralde, Elisa; Weichenrieder, Oliver

doi:10.1128/mcb.01506-07

Cited by 75 publications

(124 citation statements)

References 49 publications

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“…To discriminate between these possibilities, we examined the association of DCP1a with additional components of the decapping complex (i.e., EDC4, DCP2, EDC3, and DDX6/RCK). These all coimmunoprecipitated with DCP1a, as reported before (8,(11)(12)(13)(14) (Fig. 3 A-D, lanes 9).…”

Section: Trimerization Is Required For Dcp1a To Interact With Dcp2 Anmentioning

confidence: 71%

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DCP1 forms asymmetric trimers to assemble into active mRNA decapping complexes in metazoa

Tritschler

Braun

Motz

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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DCP1 stimulates the decapping enzyme DCP2, which removes the mRNA 5 cap structure committing mRNAs to degradation. In multicellular eukaryotes, DCP1-DCP2 interaction is stabilized by additional proteins, including EDC4. However, most information on DCP2 activation stems from studies in S. cerevisiae, which lacks EDC4. Furthermore, DCP1 orthologs from multicellular eukaryotes have a C-terminal extension, absent in fungi. Here, we show that in metazoa, a conserved DCP1 C-terminal domain drives DCP1 trimerization. Crystal structures of the DCP1-trimerization domain reveal an antiparallel assembly comprised of three kinked ␣-helices. Trimerization is required for DCP1 to be incorporated into active decapping complexes and for efficient mRNA decapping in vivo. Our results reveal an unexpected connectivity and complexity of the mRNA decapping network in multicellular eukaryotes, which likely enhances opportunities for regulating mRNA degradation.DCP2 ͉ miRNAs ͉ P-bodies ͉ EDC4 ͉ Ge-1 I n eukaryotes, removal of the mRNA 5Ј cap structure is catalyzed by the decapping enzyme DCP2 (1, 2); to be fully active and/or stable, DCP2 requires additional proteins (1, 2). Yeast DCP2 interacts directly with DCP1 and this interaction is required for decapping in vivo and in vitro (3-7). In humans, the DCP2-DCP1 interaction requires additional proteins, which together assemble into multimeric decapping complexes that also include the enhancers of decapping 3 and 4 (EDC3 and ECD4), and the DEAD-box protein DDX6/RCK (8, 9).DCP2 is highly conserved and most information on DCP2 activation stems mainly from studies in S. cerevisiae and S. pombe (3-7). Fungi, however, lack EDC4 as well as many extensions and additional domains present in decapping activators of metazoan orthologs (8-10). For example, all eukaryotic DCP1 proteins contain an N-terminal EVH1 domain (3, 5, 6); however, DCP1 orthologs from metazoa and plants also have a proline-rich C-terminal extension (9, 10). The sequence of this extension is not conserved except for a 14-residue short motif (motif I, MI) conserved in metazoa with the exception of C. elegans (Fig. 1A and Fig. S1) and a C-terminal domain conserved in plants and metazoa ( Fig. 1 A, referred to as TD).The DCP1 C-terminal domain is predicted to adopt an ␣-helical conformation. In this work, we show that this domain trimerizes in an asymmetric fashion. We solved the crystal structure of the trimerized domain for both human and D. melanogaster DCP1 and show that the trimer adopts an unprecedented fold, with no current similarities in the protein database. We further show that DCP1 trimerization is required for the assembly of active decapping complexes and for mRNA decapping in vivo. The conservation of structurally critical residues indicates that this domain adopts a similar fold in DCP1 orthologs of other multicellular eukaryotes. Consequently, within mRNA decapping complexes in these organisms, the stoichiometry of the protein components is likely more complex than previously thought. Results and DiscussionCry...

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Section: Trimerization Is Required For Dcp1a To Interact With Dcp2 Anmentioning

confidence: 71%

“…3 A-D, lanes 9). DCP1-DCP2 association is likely stabilized by endogeneous EDC4 or other components (8,9,12,14). We found DCP1a trimerization-defective mutants 1, 2, and 3 and DCP1a-⌬TD were strongly impaired in the interaction with EDC4 and DCP2 (Fig.…”

Section: Trimerization Is Required For Dcp1a To Interact With Dcp2 Anmentioning

confidence: 76%

DCP1 forms asymmetric trimers to assemble into active mRNA decapping complexes in metazoa

Tritschler

Braun

Motz

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…As a positive control, Me31B coimmunoprecipitated with HA-EDC3 as reported before ( Fig. 4B; Tritschler et al 2007). …”

Section: The C-terminal Region Of D Melanogaster Ge-1 Remains Monomementioning

confidence: 97%

The C-terminal region of Ge-1 presents conserved structural features required for P-body localization

Jinek

Eulálio²,

Lingel³

et al. 2008

RNA

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The removal of the 59 cap structure by the DCP1-DCP2 decapping complex irreversibly commits eukaryotic mRNAs to degradation. In human cells, the interaction between DCP1 and DCP2 is bridged by the Ge-1 protein. Ge-1 contains an Nterminal WD40-repeat domain connected by a low-complexity region to a conserved C-terminal domain. It was reported that the C-terminal domain interacts with DCP2 and mediates Ge-1 oligomerization and P-body localization. To understand the molecular basis for these functions, we determined the three-dimensional crystal structure of the most conserved region of the Drosophila melanogaster Ge-1 C-terminal domain. The region adopts an all a-helical fold related to ARM-and HEAT-repeat proteins. Using structure-based mutants we identified an invariant surface residue affecting P-body localization. The conservation of critical surface and structural residues suggests that the C-terminal region adopts a similar fold with conserved functions in all members of the Ge-1 protein family.

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“…Lsm12 includes t-RNA and methyltransferase domains (Albrecht & Lengauer, 2004), and Lsm13, Lsm14 and Lsm15 all contain a central DFDF-x(7)-F domain (Albrecht & Lengauer, 2004;Anantharaman & Aravind, 2004). Lsm16 features a remarkably disrupted Lsm variant (lacking both the N-terminal -helix and a complete 4 strand) in addition to FDF and YjeF-N domains (Albrecht & Lengauer, 2004;Tritschler et al, 2007). This protein is suggested to be dimeric in solution (Ling et al, 2008).…”

Section: Phylogeny Of Lsm Protein Sequencesmentioning

confidence: 99%