Abstract
Previous studies indicated that the P-body components, CGH-1 and EDC-3 may play a crucial role in the regulation of lifespan in Caenorhabditis elegans. Homo sapiens DDX6 or Saccharomyces cerevisiae Dhh1p (CGH-1 in C. elegans) could form complexes with EDC3 (Edc3p in yeast), respectively, which is significant for translation inhibition and mRNA decay. However, it is currently unclear how CGH-1 can be recognized by EDC-3 in C. elegans. Here, we provided structural and biochemical insights into the interaction between CGH-1 and EDC-3. Combined with homology modeling, mutation, and ITC assays, we uncovered an interface between CGH-1 RecA2 domain and EDC-3 FDF-FEK. Additionally, GST-pulldown and co-localization experiments confirmed the interaction between CGH-1 and EDC-3 in vitro and in vivo. We also analyzed PATR-1-binding interface on CGH-1 RecA2 by ITC assays. Moreover, we unveiled the similarity and differences of the binding mode between EDC-3 and CAR-1 or PATR-1. Taken together, these findings provide insights into the recognition of DEAD-box protein CGH-1 by EDC-3 FDF-FEK motif, suggesting important functional implications.
Highlights
Previous studies indicated that the P-body components, CGH-1 and enhancer of decapping-3 (EDC-3) may play a crucial role in the regulation of lifespan in Caenorhabditis elegans
Based on homology modeling [we are not able to crystalize the complex of CGH-1248–420/EDC-3235–271], mutation and biochemical analyses, we demonstrated that EDC-3 FDF-FEK is anchored to CGH-1 RecA2 domain through a conserved hydrophobic surface
CGH-1 is similar to its orthologs DDX6/Dhh1p, which contains two RecA-like domains connected by a short liker (Fig. 1A)
Summary
Previous studies indicated that the P-body components, CGH-1 and EDC-3 may play a crucial role in the regulation of lifespan in Caenorhabditis elegans. Homo sapiens DDX6 or Saccharomyces cerevisiae Dhh1p (CGH-1 in C. elegans) could form complexes with EDC3 (Edc3p in yeast), respectively, which is significant for translation inhibition and mRNA decay. We unveiled the similarity and differences of the binding mode between EDC-3 and CAR-1 or PATR-1 Taken together, these findings provide insights into the recognition of DEAD-box protein CGH-1 by EDC-3 FDF-FEK motif, suggesting important functional implications. The human DEAD (Asp-Glu-Ala-Asp) box DDX6 and its orthologs in X. laevis (Xp54), D. melanogaster (Me31B), C. elegans (CGH-1), and S. cerevisiae (Dhh1p) (Fig. 1A,B) play a critical role in posttranscriptional gene regulation by mediating both translational repression and mRNA decapping (reviewed in5,6). The corresponding mutant protein lacks 202 amino acids, including the conserved FDF domain of EDC-315 This line of evidence implies that EDC-3 FDF mediated recruitment of CGH-1 might be involved in the regulation of lifespan in C. elegans
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