Dynamics of re-constitution of the human nuclear proteome after cell division is regulated by NLS-adjacent phosphorylation

Gergely Róona, Máatée Borsos, Jonathan J. Ellis, Ahmed M. Mehdi, Mary Christie, Zsuzsanna Köornyei, Máatée Neubrandt, Judit Tóoth, Zoltáan Bozóoky, Láaszlóo Buday, Emília Madaráasz, Mikael Bodáen, Bostjan Kobe, Beáata G. Véertessy

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Phosphorylation by the cyclin-dependent kinase 1 (Cdk1) adjacent to nuclear localization signals (NLSs) is an important mechanism of regulation of nucleocytoplasmic transport. However, no systematic survey has yet been performed in human cells to analyze this regulatory process, and the corresponding cell-cycle dynamics have not yet been investigated. Here, we focused on the human proteome and found that numerous proteins, previously not identifi ed in this context, are associated with Cdk1-dependent phosphorylation sites adjacent to their NLSs. Interestingly, these proteins are involved in key regulatory events of DNA repair, epigenetics, or RNA editing and splicing. This finding indicates that cell-cycle dependent events of genome editing and gene expression profiling may be controlled by nucleocytoplasmic trafficking. For in-depth investigations, we selected a number of these proteins and analyzed how point mutations, expected to modify the phosphorylation ability of the NLS segments, perturb nucleocytoplasmic localization. In each case, we found that mutations mimicking hyper-phosphorylation abolish nuclear import processes. To understand the mechanism underlying these phenomena, we performed a video microscopy-based kinetic analysis to obtain information on cell-cycle dynamics on a model protein, dUTPase. We show that the NLS-adjacent phosphorylation by Cdk1 of human dUTPase, an enzyme essential for genomic integrity, results in dynamic cell cycle-dependent distribution of the protein. Non-phosphorylatable mutants have drastically altered protein re-import characteristics into the nucleus during the G1 phase. Our results suggest a dynamic Cdk1-driven mechanism of regulation of the nuclear proteome composition during the cell cycle.

Original languageEnglish
Pages (from-to)3551-3564
Number of pages14
JournalCell Cycle
Volume13
Issue number22
DOIs
Publication statusPublished - Nov 15 2014

    Fingerprint

Keywords

  • Cell cycle
  • DUTPase
  • Importin
  • Phosphorylation
  • Trafficking

ASJC Scopus subject areas

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

Róona, G., Borsos, M., Ellis, J. J., Mehdi, A. M., Christie, M., Köornyei, Z., Neubrandt, M., Tóoth, J., Bozóoky, Z., Buday, L., Madaráasz, E., Bodáen, M., Kobe, B., & Véertessy, B. G. (2014). Dynamics of re-constitution of the human nuclear proteome after cell division is regulated by NLS-adjacent phosphorylation. Cell Cycle, 13(22), 3551-3564. https://doi.org/10.4161/15384101.2014.960740