Insight into the interaction between the RNA helicase CGH-1 and EDC-3 and its implications

• Published in: Scientific reports 2021-10, Vol.11 (1), p.20359-20359, Article 20359
• Main Authors: Zhang, Yong, Wang, Ke, Yang, Kanglong, Shi, Yunyu, Hong, Jingjun
• Format: Article
• Language: English
• Subjects: 631/1647; 631/45; Animals; Caenorhabditis elegans - embryology; Caenorhabditis elegans - genetics; Caenorhabditis elegans Proteins - chemistry; Caenorhabditis elegans Proteins - genetics; Cloning; DNA helicase; E coli; Enzymes; Homology; Humanities and Social Sciences; Humans; Laboratories; Life sciences; Life span; Localization; mRNA turnover; multidisciplinary; Mutation; Peptides; Plasmids; Protein Binding; Protein expression; Proteins; RNA helicase; RNA Helicases - chemistry; RNA Helicases - genetics; RNA Nucleotidyltransferases - chemistry; RNA Nucleotidyltransferases - genetics; Saccharomyces cerevisiae - enzymology; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae Proteins - chemistry; Saccharomyces cerevisiae Proteins - genetics; Science; Science (multidisciplinary); Worms; Yeasts
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-021-99919-0

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.