@article{8394235c98024c0b8adeb883ffe8a604,
title = "The Translation Inhibitor Rocaglamide Targets a Bimolecular Cavity between eIF4A and Polypurine RNA",
abstract = "A class of translation inhibitors, exemplified by the natural product rocaglamide A (RocA), isolated from Aglaia genus plants, exhibits antitumor activity by clamping eukaryotic translation initiation factor 4A (eIF4A) onto polypurine sequences in mRNAs. This unusual inhibitory mechanism raises the question of how the drug imposes sequence selectivity onto a general translation factor. Here, we determined the crystal structure of the human eIF4A1⋅ATP analog⋅RocA⋅polypurine RNA complex. RocA targets the “bi-molecular cavity” formed characteristically by eIF4A1 and a sharply bent pair of consecutive purines in the RNA. Natural amino acid substitutions found in Aglaia eIF4As changed the cavity shape, leading to RocA resistance. This study provides an example of an RNA-sequence-selective interfacial inhibitor fitting into the space shaped cooperatively by protein and RNA with specific sequences. Iwasaki et al. resolve the structure of the human eIF4A1⋅AMPPNP⋅RocA⋅polypurine RNA complex and identify the natural amino acid substitutions in Aglaia eIF4As. These explain the molecular basis of RNA sequence selectivity provided by RocA and the resistance mechanism of the Aglaia plant, a natural source of RocA.",
keywords = "FMO, Ribo-Seq, Rocaglamide, X-ray, crystal structure, eIF4A, evolution, ribosome profiling, sequence specificity, translation",
author = "Shintaro Iwasaki and Wakana Iwasaki and Mari Takahashi and Ayako Sakamoto and Chiduru Watanabe and Yuichi Shichino and Floor, {Stephen N.} and Koichi Fujiwara and Mari Mito and Kosuke Dodo and Mikiko Sodeoka and Hiroaki Imataka and Teruki Honma and Kaori Fukuzawa and Takuhiro Ito and Ingolia, {Nicholas T.}",
note = "Funding Information: We thank Zuriah Meacham, Chris Meacham, and The University and Jepson Herbaria (University of California, Berkeley) for the reference of Aglaia odorata , the staff of the beamline BL41XU at the SPring-8 for their support during data collection, and Drs. Yoshio Okiyama and Tatsuya Nakano for helping with RNA fragmentation in the FMO calculation. We are also grateful to all the members of the Ingolia, Lareau, and Iwasaki laboratories for faithful discussions, technical help, and critical reading of the manuscript. N.T.I. was supported by the Damon Runyon Cancer Research Foundation (grant DRR-37-15 ), the Searle Scholars Program (grant 11-SSP-229 ), and the National Institute of General Medical Sciences of the NIH (grant P50GM102706 ). T.I. was supported by the Japan Society for the Promotion of Science (JSPS) (Grants-in-Aid for Scientific Research on Innovative Areas “nascent chain biology” JP15H01548 and JP17H05677 and Grant-in-Aid for Scientific Research [B] JP16H04756 ), RIKEN (the Aging Project and the RIKEN Pioneering Project “Dynamic Structural Biology”), and the Takeda Science Foundation . S.I. was supported by the JSPS (Grant-in-Aid for Scientific Research on Innovative Areas “nascent chain biology” JP17H05679 and Grant-in-Aid for Young Scientists [A] JP17H04998 ), and RIKEN (RIKEN Pioneering Projects “Cellular Evolution,” the All RIKEN Research Project “Disease and Epigenome” and the Aging Project). This work was also supported by AMED (the Platform Project for Supporting Drug Discovery, Life Science Research, Basis for Supporting Innovative Drug Discovery and Life Science Research [BINDS] JP18am0101082 and JP18am0101113 ), and AMED-CREST (grant JP18gm0710004 ). S.N.F. was a Howard Hughes Medical Institute fellow of the Helen Hay Whitney Foundation . S.I. was a recipient of Human Frontier Science Program long-term fellowship. DNA libraries were sequenced by the Vincent J. Coates Genomics Sequencing Laboratory at the University of California, Berkeley, which is supported by the NIH (S10 instrumentation grants S10RR029668 , S10RR027303 , and OD018174 ). Computations were supported by Manabu Ishii, Itoshi Nikaido, and the Bioinformatics Analysis Environment Service on the RIKEN Cloud at the RIKEN Advance Center. Crystal data acquisition was performed under the approval of the Japan Synchrotron Radiation Research Institute (proposals 2017A2581 and 2017B2727 ). Funds for the 900 MHz NMR spectrometer were provided by the NIH (grant GM68933 ). Funding Information: We thank Zuriah Meacham, Chris Meacham, and The University and Jepson Herbaria (University of California, Berkeley) for the reference of Aglaia odorata, the staff of the beamline BL41XU at the SPring-8 for their support during data collection, and Drs. Yoshio Okiyama and Tatsuya Nakano for helping with RNA fragmentation in the FMO calculation. We are also grateful to all the members of the Ingolia, Lareau, and Iwasaki laboratories for faithful discussions, technical help, and critical reading of the manuscript. N.T.I. was supported by the Damon Runyon Cancer Research Foundation (grant DRR-37-15), the Searle Scholars Program (grant 11-SSP-229), and the National Institute of General Medical Sciences of the NIH (grant P50GM102706). T.I. was supported by the Japan Society for the Promotion of Science (JSPS) (Grants-in-Aid for Scientific Research on Innovative Areas “nascent chain biology” JP15H01548 and JP17H05677 and Grant-in-Aid for Scientific Research [B] JP16H04756), RIKEN (the Aging Project and the RIKEN Pioneering Project “Dynamic Structural Biology”), and the Takeda Science Foundation. S.I. was supported by the JSPS (Grant-in-Aid for Scientific Research on Innovative Areas “nascent chain biology” JP17H05679 and Grant-in-Aid for Young Scientists [A] JP17H04998), and RIKEN (RIKEN Pioneering Projects “Cellular Evolution,” the All RIKEN Research Project “Disease and Epigenome” and the Aging Project). This work was also supported by AMED (the Platform Project for Supporting Drug Discovery, Life Science Research, Basis for Supporting Innovative Drug Discovery and Life Science Research [BINDS] JP18am0101082 and JP18am0101113), and AMED-CREST (grant JP18gm0710004). S.N.F. was a Howard Hughes Medical Institute fellow of the Helen Hay Whitney Foundation. S.I. was a recipient of Human Frontier Science Program long-term fellowship. DNA libraries were sequenced by the Vincent J. Coates Genomics Sequencing Laboratory at the University of California, Berkeley, which is supported by the NIH (S10 instrumentation grants S10RR029668, S10RR027303, and OD018174). Computations were supported by Manabu Ishii, Itoshi Nikaido, and the Bioinformatics Analysis Environment Service on the RIKEN Cloud at the RIKEN Advance Center. Crystal data acquisition was performed under the approval of the Japan Synchrotron Radiation Research Institute (proposals 2017A2581 and 2017B2727). Funds for the 900 MHz NMR spectrometer were provided by the NIH (grant GM68933). Publisher Copyright: {\textcopyright} 2018 Elsevier Inc.",
year = "2019",
month = feb,
day = "21",
doi = "10.1016/j.molcel.2018.11.026",
language = "English",
volume = "73",
pages = "738--748.e9",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "4",
}