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Quantitative Proteomic Analysis of Enriched Nuclear Fractions from BK Polyomavirus-Infected Primary Renal Proximal Tubule Epithelial Cells
- Source :
- Journal of Proteome Research. 14:4413-4424
- Publication Year :
- 2015
- Publisher :
- American Chemical Society (ACS), 2015.
-
Abstract
- Polyomaviruses are a family of small DNA viruses that are associated with a number of severe human diseases, particularly in immunocompromised individuals. The detailed virus-host interactions during lytic polyomavirus infection are not fully understood. Here, we report the first nuclear proteomic study with BK polyomavirus (BKPyV) in a primary renal proximal tubule epithelial cell culture system using stable isotope labeling by amino acids in cell culture (SILAC) proteomic profiling coupled with liquid chromatography-tandem mass spectrometry. We demonstrated the feasibility of SILAC labeling in these primary cells and subsequently performed reciprocal labeling-infection experiments to identify proteins that are altered by BKPyV infection. Our analyses revealed specific proteins that are significantly up- or down-regulated in the infected nuclear proteome. The genes encoding many of these proteins were not identified in a previous microarray study, suggesting that differential regulation of these proteins may be independent of transcriptional control. Western blotting experiments verified the SILAC proteomic findings. Finally, pathway and network analyses indicated that the host cell DNA damage response signaling and DNA repair pathways are among the cellular processes most affected at the protein level during polyomavirus infection. Our study provides a comprehensive view of the host nuclear proteomic changes during polyomavirus lytic infection and suggests potential novel host factors required for a productive polyomavirus infection.
- Subjects :
- DNA Repair
Proteome
Transcription, Genetic
Primary Cell Culture
Biology
medicine.disease_cause
Biochemistry
Article
Kidney Tubules, Proximal
Tandem Mass Spectrometry
Stable isotope labeling by amino acids in cell culture
medicine
Humans
Gene Regulatory Networks
Gene
Cell Nucleus
Proteomic Profiling
Epithelial Cells
Molecular Sequence Annotation
General Chemistry
Molecular biology
BK virus
Cell nucleus
medicine.anatomical_structure
Gene Expression Regulation
Lytic cycle
Cell culture
BK Virus
Isotope Labeling
Host-Pathogen Interactions
Chromatography, Liquid
DNA Damage
Signal Transduction
Subjects
Details
- ISSN :
- 15353907 and 15353893
- Volume :
- 14
- Database :
- OpenAIRE
- Journal :
- Journal of Proteome Research
- Accession number :
- edsair.doi.dedup.....6873e1ff7707bce9849849b4d0b82493