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Loss of VRK1 alters the nuclear phosphoproteome in the DNA damage response to doxorubicin
Elena Navarro-Carrasco1, Aurora Campos-Díaz1, Eva Monte-Serrano1
1Molecular Mechanisms of Cancer Program, Instituto de Biología Molecular y Celular del Cáncer, Consejo Superior de Investigaciones Científicas (CSIC) - Universidad de Salamanca, E-37007, Salamanca, Spain; Instituto de Investigación Biomédica de Salamanca (IBSAL), Hospital Universitario de Salamanca, E-37007, Salamanca, Spain.
Abstract:
Dynamic chromatin remodeling requires regulatory mechanisms for its adaptation to different nuclear function, which are likely to be mediated by signaling proteins. In this context, VRK1 is a nuclear Ser-Thr kinase that regulates pathways associated with transcription, replication, recombination, and DNA repair. Therefore, VRK1 is a potential regulatory, or coordinator, molecule in these processes. In this work we studied the effect that VRK1 depletion has on the basal nuclear and chromatin phosphoproteome, and their associated pathways. VRK1 depletion caused an alteration in the pattern of the nuclear phosphoproteome, which is mainly associated with nucleoproteins, ribonucleoproteins, RNA splicing and processing. Next, it was determined the changes in proteins associated with DNA damage that was induced by doxorubicin treatment. Doxorubicin alters the nuclear phosphoproteome affecting proteins implicated in DDR, including DSB repair proteins NBN and 53BP1, cellular response to stress and chromatin organization proteins. In VRK1-depleted cells, the effect of doxorubicin on protein phosphorylation was reverted to basal levels. The nuclear phosphoproteome patterns induced by doxorubicin are altered by VRK1 depletion, and is enriched in histone modification proteins and chromatin associated proteins. These results indicate that VRK1 plays a major role in processes requiring chromatin remodeling in its adaptation to different biological contexts.
Insights
Vaccinia-related kinase 1 (VRK1) depletion alters nuclear phosphoproteome, impacting DNA damage response and chromatin remodeling. VRK1 is crucial for adapting nuclear functions during cellular stress and DNA repair.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Dynamic chromatin remodeling is essential for nuclear function and relies on signaling proteins.
- Vaccinia-related kinase 1 (VRK1) is a nuclear serine-threonine kinase involved in transcription, replication, recombination, and DNA repair.
- VRK1's role as a potential coordinator molecule in these nuclear processes warrants investigation.
Purpose of the Study:
- To investigate the impact of VRK1 depletion on the basal nuclear and chromatin phosphoproteome.
- To analyze the effect of VRK1 depletion on DNA damage-induced protein phosphorylation changes.
Main Methods:
- Depletion of VRK1 in cells.
- Phosphoproteomic analysis of nuclear and chromatin fractions.
- Treatment with doxorubicin to induce DNA damage.
- Comparative analysis of phosphoproteome patterns.
Main Results:
- VRK1 depletion altered the nuclear phosphoproteome, affecting nucleoproteins, ribonucleoproteins, and RNA splicing pathways.
- Doxorubicin treatment induced changes in DNA damage response (DDR) proteins, including NBN and 53BP1, and chromatin organization proteins.
- In VRK1-depleted cells, doxorubicin-induced phosphorylation changes were reverted to basal levels.
- VRK1 depletion altered doxorubicin-induced phosphoproteome patterns, enriching histone modification and chromatin-associated proteins.
Conclusions:
- VRK1 plays a significant role in regulating the nuclear phosphoproteome.
- VRK1 is critical for the cellular adaptation of chromatin remodeling processes during DNA damage and stress.
- VRK1 acts as a key regulator in DNA repair and chromatin organization pathways.
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