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

Roona, Gergely, Borsos, Mate, Ellis, Jonathan J., Mehdi, Ahmed M., Christie, Mary, Koornyei, Zsuzsanna, Neubrandt, Mate, Tooth, Judit, Bozooky, Zoltan, Buday, Laszlo, Madaraasz, Emilia, Bodaen, Mikael, Kobe, Bostjan and Veertessy, Beata 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. doi:10.4161/15384101.2014.960740


Author Roona, Gergely
Borsos, Mate
Ellis, Jonathan J.
Mehdi, Ahmed M.
Christie, Mary
Koornyei, Zsuzsanna
Neubrandt, Mate
Tooth, Judit
Bozooky, Zoltan
Buday, Laszlo
Madaraasz, Emilia
Bodaen, Mikael
Kobe, Bostjan
Veertessy, Beata G.
Title Dynamics of re-constitution of the human nuclear proteome after cell division is regulated by NLS-adjacent phosphorylation
Journal name Cell Cycle   Check publisher's open access policy
ISSN 1551-4005
1538-4101
Publication date 2014-11-15
Year available 2014
Sub-type Article (original research)
DOI 10.4161/15384101.2014.960740
Open Access Status
Volume 13
Issue 22
Start page 3551
End page 3564
Total pages 14
Place of publication Austin, TX United States
Publisher Landes Bioscience
Collection year 2015
Language eng
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 identified 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.
Keyword Cell cycle
DUTPase
Importin
Phosphorylation
Trafficking
dNTP
Deoxyribonucleotide triphosphate
dTTP
Deoxythymindine triphosphate
dUMP
dUTP
Deoxyuridine triphosphate
Cdc28
Cyclin-dependent protein kinase (Cdk) encoded by CDC28
Cdk1
Cyclin-dependent kinase 1
GO
Gene ontology
NLS
Nuclear localization signal
NES
Nuclear export signal
cNLS
Classical nuclear localization signal
SNP
Single nucleotide polymorphisms
SV40
Simian virus 40
UBA1
Ubiquitin-activating enzyme E1
UNG2
Human uracil-DNA glycosylase 2
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

 
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