Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin

Schneider‐Poetsch, Tilman; Ju, Jianhua; Eyler, Daniel E.; Dang, Yongjun; Bhat, Shridhar; Merrick, William C.; Green, Rachel; Shen, Ben; Liu, Jun O.

doi:10.1038/nchembio.304

Cited by 788 publications

(697 citation statements)

References 42 publications

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“…6b). Although the value for lactimidomycin was in agreement with the published data, cycloheximide appeared to have a much higher efficacy than reported for mammalian ribosomes using a footprinting technique 18 . Direct measurements of the binding affinity of cycloheximide to yeast ribosomes using isothermal titration calorimetry yielded a K d 5 0.14 6 0.05 mM, in agreement with the result of our competition studies (Extended Data Fig.…”

Section: S E-site Accessibility For Inhibitorssupporting

confidence: 88%

“…5). The structures explain previous biochemical data showing that cycloheximide and lactimidomycin compete with the binding of the tRNA CCA-end in the E-site of the large subunit 18 ( Fig. 3d).…”

Section: Research Articlesupporting

confidence: 86%

“…Lactimidomycin bears an additional lactone ring that is positioned on top of eL42 and directed towards the subunit interface. Although chemically unrelated to glutarimides, phyllanthoside makes contact with the same rRNA nucleotides and interacts with eL42 in a manner resembling the tRNA 18,20 . Homoharringtonine is a marketed drug for the treatment of chronic myeloid leukaemia 37 .…”

Section: The 60s Trna E-sitementioning

confidence: 99%

“…Lactimidomycin preferentially arrests ribosomes at the first peptide bond, whereas cycloheximide stalls ribosomes during ongoing translation [18][19][20] .…”

Section: S E-site Accessibility For Inhibitorsmentioning

confidence: 99%

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Structural basis for the inhibition of the eukaryotic ribosome

Loubresse

Prokhorova

Holtkamp

et al. 2014

Nature

450

449

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Section: S E-site Accessibility For Inhibitorssupporting

confidence: 88%

Section: Research Articlesupporting

confidence: 86%

Section: The 60s Trna E-sitementioning

confidence: 99%

“…Lactimidomycin preferentially arrests ribosomes at the first peptide bond, whereas cycloheximide stalls ribosomes during ongoing translation [18][19][20] .…”

Section: S E-site Accessibility For Inhibitorsmentioning

confidence: 99%

See 2 more Smart Citations

Structural basis for the inhibition of the eukaryotic ribosome

Loubresse

Prokhorova

Holtkamp

et al. 2014

Nature

450

449

View full text Add to dashboard Cite

“…Cycloheximide (CHX; ref. 23), bafilomycin A1 (BafA1; ref. 24), chloroquine (25), and monensin (26) were from Sigma.…”

Section: Methodsmentioning

confidence: 99%

Bispecific Antibodies and Antibody–Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs

Andreev

Thambi

Bay

et al. 2017

Molecular Cancer Therapeutics

108

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The properties of cell surface proteins targeted by antibodydrug conjugates (ADCs) have not been fully exploited; of particular importance are the rate of internalization and the route of intracellular trafficking. In this study, we compared the trafficking of HER2, which is the target of the clinically approved ADC ado-trastuzumab emtansine (T-DM1), with that of prolactin receptor (PRLR), another potential target in breast cancer. In contrast to HER2, we found that PRLR is rapidly and constitutively internalized, and traffics efficiently to lysosomes, where it is degraded. The PRLR cytoplasmic domain is necessary to promote rapid internalization and degradation, and when transferred to HER2, enhances HER2 degradation. In accordance with these findings, low levels of cell surface PRLR ($30,000 surface receptors per cell) are sufficient to mediate effective killing by PRLR ADC, whereas cell killing by HER2 ADC requires higher levels of cell surface HER2 ($10 6 surface receptors per cell). Noncovalently crosslinking HER2 to PRLR at the cell surface, using a bispecific antibody that binds to both receptors, dramatically enhances the degradation of HER2 as well as the cell killing activity of a noncompeting HER2 ADC. Furthermore, in breast cancer cells that coexpress HER2 and PRLR, a HER2xPRLR bispecific ADC kills more effectively than HER2 ADC. These results emphasize that intracellular trafficking of ADC targets is a key property for their activity and, further, that coupling an ADC target to a rapidly internalizing protein may be a useful approach to enhance internalization and cell killing activity of ADCs.

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Genome‐Wide Annotation and Quantitation of Translation by Ribosome Profiling

Ingolia

Brar

Rouskin

et al. 2013

CP Molecular Biology

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Recent studies highlight the importance of translational control in determining protein abundance, underscoring the value of measuring gene expression at the level of translation. We present a protocol for genome-wide, quantitative analysis of in vivo translation by deep sequencing. This ribosome profiling approach maps the exact positions of ribosomes on transcripts by nuclease footprinting. The nuclease-protected mRNA fragments are converted into a DNA library suitable for deep sequencing using a strategy that minimizes bias. The abundance of different footprint fragments in deep sequencing data reports on the amount of translation of a gene. Additionally, footprints reveal the exact regions of the transcriptome that are translated. To better define translated reading frames, we describe an adaptation that reveals the sites of translation initiation by pre-treating cells with harringtonine to immobilize initiating ribosomes. The protocol we describe requires 5 – 7 days to generate a completed ribosome profiling sequencing library. Sequencing and data analysis requires a further 4 – 5 days.

show abstract

Inhibition of eukaryotic translation elongation by cycloheximide and lactimidomycin

Cited by 788 publications

References 42 publications

Structural basis for the inhibition of the eukaryotic ribosome

Structural basis for the inhibition of the eukaryotic ribosome

Bispecific Antibodies and Antibody–Drug Conjugates (ADCs) Bridging HER2 and Prolactin Receptor Improve Efficacy of HER2 ADCs

Genome‐Wide Annotation and Quantitation of Translation by Ribosome Profiling

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