DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity

Tatiana N. Moiseeva, Andrew Bottrill, Gerry Melino, Nickolai A. Barlev

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Stability of proteins is largely controlled by post-translational covalent modifications. Among those, ubiquitylation plays a central role as it marks the proteins for proteasome-dependent degradation. Proteolytic activities of proteasomes are critical for execution of various cellular processes, including DNA damage signaling and repair. However, very little is known about the regulation of proteasomal activity in cells during genotoxic stress. Here we investigated post-translational modifications of the 20S proteasomal subunits upon DNA damage induced by doxorubicin. Using mass-spectrometry, we found novel sites of phosphorylation and ubiquitylation in multiple proteasome subunits upon doxorubicin treatment. Ectopic co-expression of proteasome subunits and tagged ubiquitin confirmed the presence of ubiquitylated forms of PSMA5, PSMA1, PSMA3 and PSMB5 in cells. Moreover, we demonstrated that ubiquitylation in vitro inhibited chymotrypsin-like and caspase-like activities of proteasomes. In vivo, doxorubicin increased the activity of proteasomes, paralleling with attenuation of the overall level of proteasome ubiquitylation. Collectively, our results suggest a novel mechanism whereby the proteolytic activities of proteasomes are dynamically regulated by ubiquitylation upon DNA damage.

Original languageEnglish
Pages (from-to)1338-1348
Number of pages11
JournalOncotarget
Volume4
Issue number9
Publication statusPublished - 2013

Fingerprint

Ubiquitination
Proteasome Endopeptidase Complex
DNA Damage
Doxorubicin
Post Translational Protein Processing
Protein Stability
Chymotrypsin
Caspases
Ubiquitin
DNA Repair
Mass Spectrometry
Phosphorylation

Keywords

  • DNA damage
  • Doxorubicin
  • Proteasome
  • Proteolytic activities
  • Ubiquitylation

ASJC Scopus subject areas

  • Oncology

Cite this

Moiseeva, T. N., Bottrill, A., Melino, G., & Barlev, N. A. (2013). DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity. Oncotarget, 4(9), 1338-1348.

DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity. / Moiseeva, Tatiana N.; Bottrill, Andrew; Melino, Gerry; Barlev, Nickolai A.

In: Oncotarget, Vol. 4, No. 9, 2013, p. 1338-1348.

Research output: Contribution to journalArticle

Moiseeva, TN, Bottrill, A, Melino, G & Barlev, NA 2013, 'DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity', Oncotarget, vol. 4, no. 9, pp. 1338-1348.
Moiseeva, Tatiana N. ; Bottrill, Andrew ; Melino, Gerry ; Barlev, Nickolai A. / DNA damage-induced ubiquitylation of proteasome controls its proteolytic activity. In: Oncotarget. 2013 ; Vol. 4, No. 9. pp. 1338-1348.
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