Protein synthesis is a highly conserved process in all kingdoms of life that can be broken down into four distinct phases: initiation, elongation, termination and ribosome recycling. The broad goals of this research program are to use biochemical and genomic approaches to shed light on the common and distinctive molecular features of translation elongation, termination, and recycling in bacteria and eukaryotes, and their control. Here we are particularly focused on one aspect of translational control in which ribosomal stalling triggers a cellular response leading to mRNA decay, targeted proteolysis, and ribosome recycling. In particular, we focused initially on a highly conserved stalling motif, the poly-basic peptide sequence, that is of particular relevance in eukaryotic cells where alternative polyadenylation site usage commonly leads to non-stop mRNAs. We will continue to use in vitro biochemistry and in vivo ribosome profiling to look at the molecular mechanics of this biologically important and conserved process (and other related systems). More specifically, we propose (1) to use our previously established in vitro reconstituted translation system (with S. cerevisiae components) to ask a series of questions about ribosome-based mechanisms for sensing translational perturbations, (2) to use a series of reporters in yeast to screen for novel components that contribute to these mRNA surveillance pathways and (3) to use ribosome profiling approaches to define the biologically relevant in vivo targets, their molecular features, and the factors that contribute to these important pathways. We anticipate that the synergy of these approaches will be powerful in defining biologically relevant mechanism.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37GM059425-19
Application #
9668259
Study Section
Special Emphasis Panel (NSS)
Program Officer
Reddy, Michael K
Project Start
1999-05-01
Project End
2024-08-31
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
19
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21205
Schuller, Anthony P; Zinshteyn, Boris; Enam, Syed Usman et al. (2018) Directed hydroxyl radical probing reveals Upf1 binding to the 80S ribosomal E site rRNA at the L1 stalk. Nucleic Acids Res 46:2060-2073
Schuller, Anthony P; Green, Rachel (2018) Roadblocks and resolutions in eukaryotic translation. Nat Rev Mol Cell Biol 19:526-541
Dever, Thomas E; Dinman, Jonathan D; Green, Rachel (2018) Translation Elongation and Recoding in Eukaryotes. Cold Spring Harb Perspect Biol 10:
Guydosh, Nicholas R; Green, Rachel (2017) Translation of poly(A) tails leads to precise mRNA cleavage. RNA 23:749-761
McClary, Brandon; Zinshteyn, Boris; Meyer, Mélanie et al. (2017) Inhibition of Eukaryotic Translation by the Antitumor Natural Product Agelastatin A. Cell Chem Biol 24:605-613.e5
Guydosh, Nicholas R; Kimmig, Philipp; Walter, Peter et al. (2017) Regulated Ire1-dependent mRNA decay requires no-go mRNA degradation to maintain endoplasmic reticulum homeostasis in S. pombe. Elife 6:
Schuller, Anthony P; Green, Rachel (2017) The ABC(E1)s of Ribosome Recycling and Reinitiation. Mol Cell 66:578-580
Schuller, Anthony P; Wu, Colin Chih-Chien; Dever, Thomas E et al. (2017) eIF5A Functions Globally in Translation Elongation and Termination. Mol Cell 66:194-205.e5
Zinshteyn, Boris; Green, Rachel (2016) When stop makes sense. Science 354:1106
Radhakrishnan, Aditya; Chen, Ying-Hsin; Martin, Sophie et al. (2016) The DEAD-Box Protein Dhh1p Couples mRNA Decay and Translation by Monitoring Codon Optimality. Cell 167:122-132.e9

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