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Translation termination factor eRF3 is targeted for caspase-mediated proteolytic cleavage and degradation during DNA damage-induced apoptosis

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Abstract

Polypeptide chain release factor eRF3 plays pivotal roles in translation termination and post-termination events including ribosome recycling and mRNA decay. It is not clear, however, if eRF3 is targeted for the regulation of gene expression. Here we show that DNA-damaging agents (UV and etoposide) induce the immediate cleavage and degradation of eRF3 in a caspase-dependent manner. The effect is selective since the binding partners of eRF3, eRF1 and PABP, and an unrelated control, GAPDH, were not affected. Point mutations of aspartate residues within overlapping DXXD motifs near the amino terminus of eRF3 prevented the appearance of the UV-induced cleavage product, identifying D32 as the major cleavage site. The cleavage and degradation occurred in a similar time-dependent manner to those of eIF4G, a previously established caspase-3 target involved in the inhibition of translation during apoptosis. siRNA-mediated knockdown of eRF3 led to inhibition of cellular protein synthesis, supporting the idea that the decrease in the amount of eRF3 caused by the caspase-mediated degradation contributes to the inhibition of translation during apoptosis. This is the first report showing that eRF3 could serve as a target in the regulation of gene expression.

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Abbreviations

eIF:

Eukaryotic translation initiation factor

eRF:

Eukaryotic translation releasing factor

PABP:

Polyadenylate-binding protein

siRNA:

Small interfering RNA

PMSF:

Phenylmethylsulfonyl fluoride

N-terminal:

Amino-terminal

C-terminal:

Carboxyl-terminal

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Acknowledgments

This work was supported in part by Grant-in-Aid for Scientific Research on Innovative Areas “RNA regulation” (No. 20112005) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and Grant-in-Aid for Scientific Research (B) (No. 21370080) from Japan Society for the Promotion of Science (to S H) and NIH grant AG014357 (to ESA).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan

    Yoshifumi Hashimoto, Nao Hosoda & Shin-ichi Hoshino

  2. Department of Biochemistry and Molecular Biology, Thomas Jefferson University, Philadelphia, PA, 19107, USA

    Pinaki Datta & Emad S. Alnemri

Authors
  1. Yoshifumi Hashimoto
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  2. Nao Hosoda
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  3. Pinaki Datta
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  4. Emad S. Alnemri
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  5. Shin-ichi Hoshino
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Correspondence to Shin-ichi Hoshino.

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Hashimoto, Y., Hosoda, N., Datta, P. et al. Translation termination factor eRF3 is targeted for caspase-mediated proteolytic cleavage and degradation during DNA damage-induced apoptosis. Apoptosis 17, 1287–1299 (2012). https://doi.org/10.1007/s10495-012-0765-7

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