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Induction of p16(INK4a) is the major barrier to proliferation when Epstein-Barr virus (EBV) transforms primary B cells into lymphoblastoid cell lines.
- Source :
-
PLoS pathogens [PLoS Pathog] 2013 Feb; Vol. 9 (2), pp. e1003187. Date of Electronic Publication: 2013 Feb 21. - Publication Year :
- 2013
-
Abstract
- To explore the role of p16(INK4a) as an intrinsic barrier to B cell transformation by EBV, we transformed primary B cells from an individual homozygous for a deletion in the CDKN2A locus encoding p16(INK4a) and p14(ARF). Using recombinant EBV-BAC viruses expressing conditional EBNA3C (3CHT), we developed a system that allows inactivation of EBNA3C in lymphoblastoid cell lines (LCLs) lacking active p16(INK4a) protein but expressing a functional 14(ARF)-fusion protein (p14/p16). The INK4a locus is epigenetically repressed by EBNA3C--in cooperation with EBNA3A--despite the absence of functional p16(INK4a). Although inactivation of EBNA3C in LCLs from normal B cells leads to an increase in p16(INK4a) and growth arrest, EBNA3C inactivation in the p16(INK4a)-null LCLs has no impact on the rate of proliferation, establishing that the repression of INK4a is a major function of EBNA3C in EBV-driven LCL proliferation. This conditional LCL system allowed us to use microarray analysis to identify and confirm genes regulated specifically by EBNA3C, independently of proliferation changes modulated by the p16(INK4a)-Rb-E2F axis. Infections of normal primary B cells with recombinant EBV-BAC virus from which EBNA3C is deleted or with 3CHT EBV in the absence of activating ligand 4-hydroxytamoxifen, revealed that EBNA3C is necessary to overcome an EBV-driven increase in p16(INK4a) expression and concomitant block to proliferation 2-4 weeks post-infection. If cells are p16(INK4a)-null, functional EBNA3C is dispensable for the outgrowth of LCLs.
- Subjects :
- Antigens, Viral genetics
Antigens, Viral metabolism
Cell Line
Cell Proliferation
Cell Survival
Cyclin-Dependent Kinase Inhibitor p16 metabolism
Epstein-Barr Virus Infections immunology
Epstein-Barr Virus Infections virology
Epstein-Barr Virus Nuclear Antigens genetics
Epstein-Barr Virus Nuclear Antigens metabolism
Genetic Loci
Herpesvirus 4, Human immunology
Humans
Oligonucleotide Array Sequence Analysis
Phosphorylation
Primary Cell Culture
Virus Latency
B-Lymphocytes virology
Cyclin-Dependent Kinase Inhibitor p16 genetics
Epigenetic Repression genetics
Herpesvirus 4, Human physiology
Lymphocyte Activation
Subjects
Details
- Language :
- English
- ISSN :
- 1553-7374
- Volume :
- 9
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- PLoS pathogens
- Publication Type :
- Academic Journal
- Accession number :
- 23436997
- Full Text :
- https://doi.org/10.1371/journal.ppat.1003187