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Cell Chem Biol. 2018 Dec 20;25(12):1506-1518.e13. doi: 10.1016/j.chembiol.2018.09.010. Epub 2018 Oct 11.

Tetracyclines Modify Translation by Targeting Key Human rRNA Substructures.

Author information

1
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
2
The Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
3
Centre for Integrative Biology (CBI), Department of Integrated Structural Biology, IGBMC (Institute of Genetics and of Molecular and Cellular Biology), 1 Rue Laurent Fries, Illkirch 67404, France; Centre National de la Recherche Scientifique (CNRS) UMR 7104, Illkirch, France; Institut National de la Santé et de la Recherche Médicale (INSERM) U964, Illkirch, France; Université de Strasbourg, Strasbourg, France.
4
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA. Electronic address: myers@chemistry.harvard.edu.

Abstract

Apart from their antimicrobial properties, tetracyclines demonstrate clinically validated effects in the amelioration of pathological inflammation and human cancer. Delineation of the target(s) and mechanism(s) responsible for these effects, however, has remained elusive. Here, employing quantitative mass spectrometry-based proteomics, we identified human 80S ribosomes as targets of the tetracyclines Col-3 and doxycycline. We then developed in-cell click selective crosslinking with RNA sequence profiling (icCL-seq) to map binding sites for these tetracyclines on key human rRNA substructures at nucleotide resolution. Importantly, we found that structurally and phenotypically variant tetracycline analogs could chemically discriminate these rRNA binding sites. We also found that tetracyclines both subtly modify human ribosomal translation and selectively activate the cellular integrated stress response (ISR). Together, the data reveal that targeting of specific rRNA substructures, activation of the ISR, and inhibition of translation are correlated with the anti-proliferative properties of tetracyclines in human cancer cell lines.

KEYWORDS:

RNA-seq; chemoproteomics; diazirine; photocrosslinking; ribosome; target identification; tetracyclines; translation

PMID:
30318461
PMCID:
PMC6309532
[Available on 2019-12-20]
DOI:
10.1016/j.chembiol.2018.09.010
[Indexed for MEDLINE]

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