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Hipp binds to the carboxyl-terminal domain of eIF4A, locks it in a closed conformation, and inhibits its RNA binding. The dependencies of mRNAs for eIF4A during initiation is contingent on the degree of secondary structure within their 5\u2032 leader region. Interest in targeting eIF4A therapeutically in cancer and viral-infected settings stems from the dependencies that certain cellular (e.g. pro-oncogenic, pro-survival) and viral mRNAs show towards eIF4A. Using a CRISPR\/Cas9-based variomics screen, we identify functional EIF4A1 Hipp-resistant alleles, which in turn allowed us to link the translation-inhibitory and cytotoxic properties of Hipp to eIF4A1 target engagement. Genome-wide translational profiling in the absence or presence of Hipp were undertaken and our validation studies provided insight into the structure-activity relationships of eIF4A-dependent mRNAs. We find that mRNA 5\u2032 leader length, overall secondary structure and cytosine content are defining features of Hipp-dependent mRNAs.<\/jats:p>","DOI":"10.1093\/nar\/gkaa662","type":"journal-article","created":{"date-parts":[[2020,8,4]],"date-time":"2020-08-04T19:28:44Z","timestamp":1596569324000},"page":"9521-9537","source":"Crossref","is-referenced-by-count":39,"title":["Identification and characterization of hippuristanol-resistant mutants reveals eIF4A1 dependencies within mRNA 5\u2032 leader regions"],"prefix":"10.1093","volume":"48","author":[{"given":"Jutta","family":"Steinberger","sequence":"first","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leo","family":"Shen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"J.\u00a0Kiniry","sequence":"additional","affiliation":[{"name":"School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sai Kiran","family":"Naineni","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Regina","family":"Cencic","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehdi","family":"Amiri","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah A E","family":"Aboushawareb","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Chu","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rayelle Itoua","family":"Ma\u00efga","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brahm\u00a0J","family":"Yachnin","sequence":"additional","affiliation":[{"name":"Department of Chemistry & Chemical Biology & the Institute for Quantitative Biomedicine, Rutgers The State University of New Jersey, Piscataway\u00a008854, NJ"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francis","family":"Robert","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nahum","family":"Sonenberg","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"},{"name":"Rosalind and Morris Goodman Cancer Research Center, McGill University, Montreal H3A 1A3, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavel V","family":"Baranov","sequence":"additional","affiliation":[{"name":"School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland"},{"name":"Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1963-6466","authenticated-orcid":false,"given":"Jerry","family":"Pelletier","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, McGill University, Montreal H3G 1Y6, Canada"},{"name":"Rosalind and Morris Goodman Cancer Research Center, McGill University, Montreal H3A 1A3, Canada"},{"name":"Department of Oncology,\u00a0McGill University, Montreal H3G 1Y6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2020,8,7]]},"reference":[{"key":"2020092420045321600_B1","doi-asserted-by":"crossref","first-page":"577","DOI":"10.1016\/j.celrep.2013.01.019","article-title":"A functional variomics tool for discovering drug-resistance genes and drug targets","volume":"3","author":"Huang","year":"2013","journal-title":"Cell Rep."},{"key":"2020092420045321600_B2","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1074\/jbc.M114.618140","article-title":"Variomics screen identifies the re-entrant loop of the calcium-activated chloride channel ANO1 that facilitates channel activation","volume":"290","author":"Bill","year":"2015","journal-title":"J. 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