ATPase activity of the DEAD-box protein Dhh1 controls processing body formation

Elife. 2016 Oct 3;5:e18746. doi: 10.7554/eLife.18746.

Abstract

Translational repression and mRNA degradation are critical mechanisms of posttranscriptional gene regulation that help cells respond to internal and external cues. In response to certain stress conditions, many mRNA decay factors are enriched in processing bodies (PBs), cellular structures involved in degradation and/or storage of mRNAs. Yet, how cells regulate assembly and disassembly of PBs remains poorly understood. Here, we show that in budding yeast, mutations in the DEAD-box ATPase Dhh1 that prevent ATP hydrolysis, or that affect the interaction between Dhh1 and Not1, the central scaffold of the CCR4-NOT complex and an activator of the Dhh1 ATPase, prevent PB disassembly in vivo. Intriguingly, this process can be recapitulated in vitro, since recombinant Dhh1 and RNA, in the presence of ATP, phase-separate into liquid droplets that rapidly dissolve upon addition of Not1. Our results identify the ATPase activity of Dhh1 as a critical regulator of PB formation.

Keywords: DEAD box ATPases; RNA turnover; S. cerevisiae; biochemistry; cell biology; processing bodies.

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism*
  • Cell Cycle Proteins / metabolism
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / metabolism*
  • Macromolecular Substances / metabolism*
  • Mutant Proteins / genetics
  • Mutant Proteins / metabolism
  • RNA, Messenger / metabolism
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Transcription Factors / metabolism

Substances

  • CDC39 protein, S cerevisiae
  • Cell Cycle Proteins
  • Macromolecular Substances
  • Mutant Proteins
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Adenosine Triphosphatases
  • DHH1 protein, S cerevisiae
  • DEAD-box RNA Helicases