DEAD-box RNA helicase Belle posttranscriptionally promotes gene expression in an ATPase activity-dependent manner

Susan E. Liao, Suresh K. Kandasamy, Li Zhu, Ryuya Fukunaga

Research output: Contribution to journalArticle

Abstract

Drosophila Belle (human ortholog DDX3) is a conserved DEAD-box RNA helicase implicated in regulating gene expression. However, the molecular mechanisms by which Belle/DDX3 regulates gene expression are poorly understood. Here we performed systematic mutational analysis to determine the contributions of conserved motifs within Belle to its in vivo function. We found that Belle RNA-binding and RNA-unwinding activities and intrinsically disordered regions (IDRs) are required for Belle in vivo function. Expression of Belle ATPase mutants that cannot bind, hydrolyze, or release ATP resulted in dominant toxic phenotypes. Mechanistically, we discovered that Belle up-regulates reporter protein level when tethered to reporter mRNA, without corresponding changes at the mRNA level, indicating that Belle promotes translation of mRNA that it binds. Belle ATPase activity and amino-terminal IDR were required for this translational promotion activity. We also found that ectopic ovary expression of dominant Belle ATPase mutants decreases levels of cyclin proteins, including Cyclin B, without corresponding changes in their mRNA levels. Finally, we found that Belle binds endogenous cyclin B mRNA. We propose that Belle promotes translation of specific target mRNAs, including cyclin B mRNA, in an ATPase activity-dependent manner.

Original languageEnglish (US)
Pages (from-to)825-839
Number of pages15
JournalRNA
Volume27
Issue number5
DOIs
StatePublished - Jan 1 2019

Fingerprint

DEAD-box RNA Helicases
Adenosine Triphosphatases
Cyclin B
Gene Expression
Messenger RNA
RNA
Cyclins
Poisons
Protein Biosynthesis
Drosophila
Ovary
Proteins
Up-Regulation
Adenosine Triphosphate
Phenotype

Keywords

  • ATPase
  • DEAD-box
  • Drosophila
  • Helicase
  • Posttranscriptional gene regulation
  • RNA-binding

ASJC Scopus subject areas

  • Molecular Biology

Cite this

DEAD-box RNA helicase Belle posttranscriptionally promotes gene expression in an ATPase activity-dependent manner. / Liao, Susan E.; Kandasamy, Suresh K.; Zhu, Li; Fukunaga, Ryuya.

In: RNA, Vol. 27, No. 5, 01.01.2019, p. 825-839.

Research output: Contribution to journalArticle

Liao, Susan E. ; Kandasamy, Suresh K. ; Zhu, Li ; Fukunaga, Ryuya. / DEAD-box RNA helicase Belle posttranscriptionally promotes gene expression in an ATPase activity-dependent manner. In: RNA. 2019 ; Vol. 27, No. 5. pp. 825-839.
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