Recruitment of the RNA helicase RHAU to stress granules via a unique RNA-binding domain.
In response to environmental stress, the translation machinery of cells is reprogrammed. The majority of actively translated mRNAs are released from polysomes and driven to specific cytoplasmic foci called stress granules (SGs) where dynamic changes in protein-RNA interaction determine the subsequent fate of mRNAs. Here we show that the DEAH box RNA helicase RHAU is a novel SG-associated protein. Although RHAU protein was originally identified as an AU-rich element-associated protein involved in urokinase-type plasminogen activator mRNA decay, it was not clear whether RHAU could directly interact with RNA. We have demonstrated that RHAU physically interacts with RNA in vitro and in vivo through a newly identified N-terminal RNA-binding domain, which was found to be both essential and sufficient for RHAU localization in SGs. We have also shown that the ATPase activity of RHAU plays a role in the RNA interaction and in the regulation of protein retention in SGs. Thus, our results show that RHAU is the fourth RNA helicase detected in SGs, after rck/p54, DDX3, and eIF4A, and that its association with SGs is dynamic and mediated by an RHAU-specific RNA-binding domain.
Friedrich Miescher Institute for Biomedical Research, Novartis Research Foundation, Maulbeerstrasse 66, 4058 Basel, Switzerland.
Katerina
K
Chalupníková
Simon
S
Lattmann
Nives
N
Selak
Fumiko
F
Iwamoto
Yukio
Y
Fujiki
Yoshikuni
Y
Nagamine
D000251:Adenosine Triphosphatases / Q000737:chemistry; D000595:Amino Acid Sequence; D003432:Cross-Linking Reagents / Q000494:pharmacology; D053487:DEAD-box RNA Helicases / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D039601:Eukaryotic Initiation Factor-4A / Q000737:chemistry; D036681:Fluorescence Recovery After Photobleaching; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008856:Microscopy, Fluorescence / Q000379:methods; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D020365:RNA Helicases / Q000737:chemistry*; D017386:Sequence Homology, Amino Acid
D003432:Cross-Linking Reagents; D039601:Eukaryotic Initiation Factor-4A; D000251:Adenosine Triphosphatases; C466988:DHX36 protein, human; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases