Proteasome activity influences UV-mediated subnuclear localization changes of NPM

PLoS One. 2013;8(3):e59096. doi: 10.1371/journal.pone.0059096. Epub 2013 Mar 12.

Abstract

UV damage activates cellular stress signaling pathways, causes DNA helix distortions and inhibits transcription by RNA polymerases I and II. In particular, the nucleolus, which is the site of RNA polymerase I transcription and ribosome biogenesis, disintegrates following UV damage. The disintegration is characterized by reorganization of the subnucleolar structures and change of localization of many nucleolar proteins. Here we have queried the basis of localization change of nucleophosmin (NPM), a nucleolar granular component protein, which is increasingly detected in the nucleoplasm following UV radiation. Using photobleaching experiments of NPM-fluorescent fusion protein in live human cells we show that NPM mobility increases after UV damage. However, we show that the increase in NPM nucleoplasmic abundance after UV is independent of UV-activated cellular stress and DNA damage signaling pathways. Unexpectedly, we find that proteasome activity affects NPM redistribution. NPM nucleolar expression was maintained when the UV-treated cells were exposed to proteasome inhibitors or when the expression of proteasome subunits was inhibited using RNAi. However, there was no evidence of increased NPM turnover in the UV damaged cells, or that ubiquitin or ubiquitin recycling affected NPM localization. These findings suggest that proteasome activity couples to nucleolar protein localizations in UV damage stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Nucleolus / metabolism
  • Cell Nucleus / metabolism*
  • Humans
  • Nuclear Proteins / metabolism*
  • Nucleophosmin
  • Proteasome Endopeptidase Complex / metabolism*
  • Proteasome Inhibitors / pharmacology
  • Protein Transport / drug effects
  • Protein Transport / radiation effects
  • RNA Processing, Post-Transcriptional / drug effects
  • RNA Processing, Post-Transcriptional / radiation effects
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism
  • Stress, Physiological
  • Transcription, Genetic / drug effects
  • Transcription, Genetic / radiation effects
  • Ubiquitin / metabolism
  • Ultraviolet Rays / adverse effects

Substances

  • NPM1 protein, human
  • Nuclear Proteins
  • Proteasome Inhibitors
  • RNA, Ribosomal
  • Ubiquitin
  • Nucleophosmin
  • Proteasome Endopeptidase Complex

Grants and funding

This work was funded by Academy of Finland (grant no. 251307), Finnish Cancer Organisations, K. Albin Johanssons stiftelse, Magnus Ehrnrooth Foundation, The Maud Kuistila Memorial Foundation, and The Ida Montin Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.