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Nuclear PKC-θ facilitates rapid transcriptional responses in human memory CD4+ T cells through p65 and H2B phosphorylation.

Li J, Hardy K, Phetsouphanh C, Tu WJ, Sutcliffe EL, McCuaig R, Sutton CR, Zafar A, Munier CM, Zaunders JJ, Xu Y, Theodoratos A, Tan A, Lim PS, Knaute T, Masch A, Zerweck J, Brezar V, Milburn PJ, Dunn J, Casarotto MG, Turner SJ, Seddiki N, Kelleher AD, Rao S - J. Cell. Sci. (2016)

Bottom Line: Memory T cells are characterized by their rapid transcriptional programs upon re-stimulation.This transcriptional memory response is facilitated by permissive chromatin, but exactly how the permissive epigenetic landscape in memory T cells integrates incoming stimulatory signals remains poorly understood.Flanked by permissive histone modifications, these PKC-enriched regions are significantly enriched with NF-κB motifs in ex vivo bulk and vaccinia-responsive human memory CD4(+) T cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Education, Science, Technology & Mathematics, University of Canberra, Canberra, Australian Capital Territory 2617, Australia Department of Microbiology & Immunology, The Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, Victoria 3010, Australia.

No MeSH data available.


Related in: MedlinePlus

PKC-θ signaling and rapid transcriptional responses in memory CD4+ T cells. (A) A schematic of the in vitro transcriptional memory Jurkat T cell model: non-stimulated (NS) Jurkat T cells were activated with PMA and Ca2+ ionophore (+P/I, denoted 1°) and then subjected to stimulus withdrawal (SW) for 9 days before re-stimulation (2°). (B) Venn diagram showing the number of genes grouped by their distinct transcriptional profiles in the Jurkat model. These profiles are for the primary-specific, activation-compliant, transcriptional-memory-responsive and secondary-specific groups. (C) Heatmap representation of inducible gene expression in naïve and memory CD4+ T cells treated with PKC-θ siRNA (siPKC) with and without PMA and Ca2+ ionophore. Gene expression normalized to GAPDH is represented as z-scores (mean, n=2). The colors of the asterisk correspond to the gene groups shown in Fig. 1B. siCtrl, control siRNA.
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JCS181248F1: PKC-θ signaling and rapid transcriptional responses in memory CD4+ T cells. (A) A schematic of the in vitro transcriptional memory Jurkat T cell model: non-stimulated (NS) Jurkat T cells were activated with PMA and Ca2+ ionophore (+P/I, denoted 1°) and then subjected to stimulus withdrawal (SW) for 9 days before re-stimulation (2°). (B) Venn diagram showing the number of genes grouped by their distinct transcriptional profiles in the Jurkat model. These profiles are for the primary-specific, activation-compliant, transcriptional-memory-responsive and secondary-specific groups. (C) Heatmap representation of inducible gene expression in naïve and memory CD4+ T cells treated with PKC-θ siRNA (siPKC) with and without PMA and Ca2+ ionophore. Gene expression normalized to GAPDH is represented as z-scores (mean, n=2). The colors of the asterisk correspond to the gene groups shown in Fig. 1B. siCtrl, control siRNA.

Mentions: To investigate the importance of PKC-θ signaling in transcriptional memory responses, we devised an in vitro transcriptional memory model in which non-stimulated Jurkat T cells were stimulated with the PKC pathway inducers PMA and Ca2+ ionophore for 4 h (denoted as the primary stimulation). This was followed by stimulus withdrawal and re-stimulation (denoted as the secondary stimulation) (Fig. 1A). Whole-transcriptomic analysis showed that a majority (but not all) stimulation-induced expression changes were reversible following stimulus removal, with expression more variable during re-stimulation (Fig. S1A). Compared to in non-stimulated cells, Gene Set Enrichment Analysis (GSEA) showed that highly expressed genes in cells subjected to stimulus withdrawal were characteristically associated with effector memory (TEM) and central memory (TCM) T cells. Similarly, more memory-cell-associated genes were upregulated in the re-stimulated (secondary) Jurkat T cells compared to cells activated by the primary stimulation (Table S1; Abbas et al., 2005, 2009; Luckey et al., 2006; Wherry et al., 2007).Fig. 1.


Nuclear PKC-θ facilitates rapid transcriptional responses in human memory CD4+ T cells through p65 and H2B phosphorylation.

Li J, Hardy K, Phetsouphanh C, Tu WJ, Sutcliffe EL, McCuaig R, Sutton CR, Zafar A, Munier CM, Zaunders JJ, Xu Y, Theodoratos A, Tan A, Lim PS, Knaute T, Masch A, Zerweck J, Brezar V, Milburn PJ, Dunn J, Casarotto MG, Turner SJ, Seddiki N, Kelleher AD, Rao S - J. Cell. Sci. (2016)

PKC-θ signaling and rapid transcriptional responses in memory CD4+ T cells. (A) A schematic of the in vitro transcriptional memory Jurkat T cell model: non-stimulated (NS) Jurkat T cells were activated with PMA and Ca2+ ionophore (+P/I, denoted 1°) and then subjected to stimulus withdrawal (SW) for 9 days before re-stimulation (2°). (B) Venn diagram showing the number of genes grouped by their distinct transcriptional profiles in the Jurkat model. These profiles are for the primary-specific, activation-compliant, transcriptional-memory-responsive and secondary-specific groups. (C) Heatmap representation of inducible gene expression in naïve and memory CD4+ T cells treated with PKC-θ siRNA (siPKC) with and without PMA and Ca2+ ionophore. Gene expression normalized to GAPDH is represented as z-scores (mean, n=2). The colors of the asterisk correspond to the gene groups shown in Fig. 1B. siCtrl, control siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4920249&req=5

JCS181248F1: PKC-θ signaling and rapid transcriptional responses in memory CD4+ T cells. (A) A schematic of the in vitro transcriptional memory Jurkat T cell model: non-stimulated (NS) Jurkat T cells were activated with PMA and Ca2+ ionophore (+P/I, denoted 1°) and then subjected to stimulus withdrawal (SW) for 9 days before re-stimulation (2°). (B) Venn diagram showing the number of genes grouped by their distinct transcriptional profiles in the Jurkat model. These profiles are for the primary-specific, activation-compliant, transcriptional-memory-responsive and secondary-specific groups. (C) Heatmap representation of inducible gene expression in naïve and memory CD4+ T cells treated with PKC-θ siRNA (siPKC) with and without PMA and Ca2+ ionophore. Gene expression normalized to GAPDH is represented as z-scores (mean, n=2). The colors of the asterisk correspond to the gene groups shown in Fig. 1B. siCtrl, control siRNA.
Mentions: To investigate the importance of PKC-θ signaling in transcriptional memory responses, we devised an in vitro transcriptional memory model in which non-stimulated Jurkat T cells were stimulated with the PKC pathway inducers PMA and Ca2+ ionophore for 4 h (denoted as the primary stimulation). This was followed by stimulus withdrawal and re-stimulation (denoted as the secondary stimulation) (Fig. 1A). Whole-transcriptomic analysis showed that a majority (but not all) stimulation-induced expression changes were reversible following stimulus removal, with expression more variable during re-stimulation (Fig. S1A). Compared to in non-stimulated cells, Gene Set Enrichment Analysis (GSEA) showed that highly expressed genes in cells subjected to stimulus withdrawal were characteristically associated with effector memory (TEM) and central memory (TCM) T cells. Similarly, more memory-cell-associated genes were upregulated in the re-stimulated (secondary) Jurkat T cells compared to cells activated by the primary stimulation (Table S1; Abbas et al., 2005, 2009; Luckey et al., 2006; Wherry et al., 2007).Fig. 1.

Bottom Line: Memory T cells are characterized by their rapid transcriptional programs upon re-stimulation.This transcriptional memory response is facilitated by permissive chromatin, but exactly how the permissive epigenetic landscape in memory T cells integrates incoming stimulatory signals remains poorly understood.Flanked by permissive histone modifications, these PKC-enriched regions are significantly enriched with NF-κB motifs in ex vivo bulk and vaccinia-responsive human memory CD4(+) T cells.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Education, Science, Technology & Mathematics, University of Canberra, Canberra, Australian Capital Territory 2617, Australia Department of Microbiology & Immunology, The Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, Victoria 3010, Australia.

No MeSH data available.


Related in: MedlinePlus