Elucidating the role of yeast Ded1, a DEAD-box RNA helicase in translation elongation

PI:  Kyle Tanner

Microbial Gene Expression Laboratory

1-year contract

DEAD-box proteins are ATP-dependent RNA binding proteins and RNA-dependent ATPases that are thought to disrupt or modify RNA secondary structures, but they can also function as ATP-dependent clamps on RNA. We used a modified PAR-CLIP technique to crosslink Ded1 to its RNA binding sites and Illuminia sequencing to identify the RNAs. We find that Ded1 interacts primarily with the ORFs of mRNAs. Moreover, by using sucrose gradients, IgG pulldowns, electromobility shift assays and enzymatic analyses of purified proteins that the ATPase activity of Ded1 is inhibited by the signal recognition particle that is involved in the transport of polypeptides into the endoplasmic reticulum during translation. Thus, Ded1 seems to regulate translating ribosomes by clamping the mRNA and pausing the ribosomes. We are interested in further elucidating the role of Ded1 and in identifying its cofactors in translation elongation.

Yeter-Alat, H., Belgareh-Touzé, N., Huvelle, E., Mokdadi, M., Banroques, J., Tanner N. K. The DEAD-box RNA helicase Ded1 from yeast is associated with the signal recognition particle (SRP), and its enzymatic activity is regulated by SRP21. BioRXiv doi: https://doi.org/10.1101/2020.11.08.373522

 

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