Cellular response to small molecules that selectively stall protein synthesis by the ribosome

Liaud, Nadège; Horlbeck, Max A.; Gilbert, Luke A.; Gjoni, Ketrin; Weissman, Jonathan S.; Cate, Jamie H.  D.

doi:10.1371/journal.pgen.1008057

Cellular response to small molecules that selectively stall protein synthesis by the ribosome

Journal Article · Fri Mar 15 00:00:00 EDT 2019 · PLoS Genetics

DOI:https://doi.org/10.1371/journal.pgen.1008057· OSTI ID:1627319

Liaud, Nadège ^[1]; ^[2]; Gilbert, Luke A. ^[3]; ^[4]; Weissman, Jonathan S. ^[2]; ^[5]

Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; DOE/OSTI
Univ. of California, San Francisco, CA (United States). Dept. of Cellular and Molecular Pharmacology; Univ. of California, San Francisco, CA (United States). Howard Hughes Medical Inst.; Univ. of California, San Francisco, CA (United States). California Inst. of Quantitative Biomedical Research
Univ. of California, San Francisco, CA (United States). Dept. of Urology; Univ. of California, San Francisco, CA (United States). Helen Diller Family Comprehensive Cancer Center
Univ. of California, Berkeley, CA (United States). Dept. of Chemistry
Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; Univ. of California, Berkeley, CA (United States). Dept. of Chemistry; Univ. of California, Berkeley, CA (United States). QB3 Inst.; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bio-imaging

Identifying small molecules that inhibit protein synthesis by selectively stalling the ribosome constitutes a new strategy for therapeutic development. Compounds that inhibit the translation of PCSK9, a major regulator of low-density lipoprotein cholesterol, have been identified that reduce LDL cholesterol in preclinical models and that affect the translation of only a few off-target proteins. Although some of these compounds hold potential for future therapeutic development, it is not known how they impact the physiology of cells or ribosome quality control pathways. Here we used a genome-wide CRISPRi screen to identify proteins and pathways that modulate cell growth in the presence of high doses of a selective PCSK9 translational inhibitor, PF-06378503 (PF8503). The two most potent genetic modifiers of cell fitness in the presence of PF8503, the ubiquitin binding protein ASCC2 and helicase ASCC3, bind to the ribosome and protect cells from toxic effects of high concentrations of the compound. Surprisingly, translation quality control proteins Pelota (PELO) and HBS1L sensitize cells to PF8503 treatment. In genetic interaction experiments, ASCC3 acts together with ASCC2, and functions downstream of HBS1L. Taken together, these results identify new connections between ribosome quality control pathways, and provide new insights into the selectivity of compounds that stall human translation that will aid the development of next-generation selective translation stalling compounds to treat disease.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1627319

Journal Information:: PLoS Genetics, Journal Name: PLoS Genetics Journal Issue: 3 Vol. 15; ISSN 1553-7404

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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