Analysis of Dom34 and its function in No-Go decay

Dario O. Passos, Meenakshi K. Doma, Christopher J. Shoemaker, Denise Muhlrad, Rachel Green, Jonathan Weissman, Julie Hollien, Roy Parker

Research output: Contribution to journalArticle

Abstract

Eukaryotic mRNAs are subject to quality control mechanisms that degrade defective mRNAs. In yeast, mRNAs with stalls in translation elongation are targeted for endonucleolytic cleavage by No-Go decay (NGD). The cleavage triggered by No-Go decay is dependent on Dom34p and Hbs1p, and Dom34 has been proposed to be the endonuclease responsible for mRNA cleavage. We created several Dom34 mutants and examined their effects on NGD in yeast. We identified mutations in several loops of the Dom34 structure that affect NGD. In contrast, mutations inactivating the proposed nuclease domain do not affect NGD in vivo. Moreover, we observed that overexpression of the Rps30a protein, a high copy suppressor of dom34Δ cold sensitivity, can restore some mRNA cleavage in a dom34Δ strain. These results identify important functional regions of Dom34 and suggest that the proposed endonuclease activity of Dom34 is not required for mRNA cleavage in NGD. We also provide evidence that the process of NGD is conserved in insect cells. On the basis of these results and the process of translation termination, we suggest a multistep model for the process of NGD.

Original languageEnglish (US)
Pages (from-to)3025-3032
Number of pages8
JournalMolecular biology of the cell
Volume20
Issue number13
DOIs
StatePublished - Jul 1 2009

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Passos, D. O., Doma, M. K., Shoemaker, C. J., Muhlrad, D., Green, R., Weissman, J., Hollien, J., & Parker, R. (2009). Analysis of Dom34 and its function in No-Go decay. Molecular biology of the cell, 20(13), 3025-3032. https://doi.org/10.1091/mbc.E09-01-0028