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

Li, Jasmine; Hardy, Kristine; Phetsouphanh, Chansavath; Tu, Wen Juan; Sutcliffe, Elissa L; McCuaig, Robert; Sutton, Christopher R; Zafar, Anjum; Munier, C. Mee Ling; Zaunders, John; Yin, Xu; Theodoratos, Angelo; Tan, Abel; Lim, Pek Siew; Knaute, Tobias; Masch, Antonia; Zerweck, Johannes; Brezar, Vedran; Milburn, Peter J.; Dunn, Jenny C.; Turner, Stephen J.; Seddiki, Nabila; Kelleher, Anthony D.; Rao, Sudha

doi:10.1242/jcs.181248

Cited by 11 publications

(22 citation statements)

References 56 publications

(64 reference statements)

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“…We have previously reported7 that nuclear NFκB family member RelA protein levels are higher in 2°-stimulated Jurkat cells, and this was confirmed by immunohistochemistry (Fig. 7a).…”

Section: Resultssupporting

confidence: 77%

“…The JTM model7 consists of four cell states: NS (non-stimulated naïve T cells), ST (PMA and ionomycin (I) stimulated), SW (stimulus withdrawal, memory-like T cells), and RS cells (re-stimulation of the memory-like T cell state) (Fig. 1a).…”

Section: Resultsmentioning

confidence: 99%

“…NFAT5 and NFATC1 show inducible expression51, with NFATC1 induced more in CD4 + memory cells than naïve cells73852. NFATC1 and NFAT5 were both induced in Jurkats, with NFATC1 induced more in RS.…”

Section: Discussionmentioning

confidence: 99%

“…Primary activated Jurkat T cells were washed three times with stimulus-free medium and re-cultured for up to six days prior to PMA/I re-stimulation as previously described7. Jurkat and HUT-78 cells were cultured in RPMI 1640 supplemented with 10% FBS and 4 mM L-glutamine.…”

Section: Methodsmentioning

confidence: 99%

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Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Hardy

Sutton

et al. 2017

Sci Rep

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Memory T cells exhibit transcriptional memory and “remember” their previous pathogenic encounter to increase transcription on re-infection. However, how this transcriptional priming response is regulated is unknown. Here we performed global FAIRE-seq profiling of chromatin accessibility in a human T cell transcriptional memory model. Primary activation induced persistent accessibility changes, and secondary activation induced secondary-specific opening of previously less accessible regions associated with enhanced expression of memory-responsive genes. Increased accessibility occurred largely in distal regulatory regions and was associated with increased histone acetylation and relative H3.3 deposition. The enhanced re-stimulation response was linked to the strength of initial PKC-induced signalling, and PKC-sensitive increases in accessibility upon initial stimulation showed higher accessibility on re-stimulation. While accessibility maintenance was associated with ETS-1, accessibility at re-stimulation-specific regions was linked to NFAT, especially in combination with ETS-1, EGR, GATA, NFκB, and NR4A. Furthermore, NFATC1 was directly regulated by ETS-1 at an enhancer region. In contrast to the factors that increased accessibility, signalling from bHLH and ZEB family members enhanced decreased accessibility upon re-stimulation. Interplay between distal regulatory elements, accessibility, and the combined action of sequence-specific transcription factors allows transcriptional memory-responsive genes to “remember” their initial environmental encounter.

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“…We have previously reported7 that nuclear NFκB family member RelA protein levels are higher in 2°-stimulated Jurkat cells, and this was confirmed by immunohistochemistry (Fig. 7a).…”

Section: Resultssupporting

confidence: 77%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Hardy

Sutton

et al. 2017

Sci Rep

Self Cite

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“…But what is the role of delocalized PKCθ in the context of Treg cell‐IS functions? In Tconv cells, upon TCR stimulation PKCθ supports NF‐κB activation and can maintain phosphorylated RelA, the p65 subunit of NF‐κB, in the nucleus . Interestingly, Treg cells were found to accumulate phosphorylated p65 at the late IS, although its functional role there remains elusive.…”

Section: Discussionmentioning

confidence: 99%

TCR signalling network organization at the immunological synapses of murine regulatory T cells

et al. 2017

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Regulatory T (Treg) cells require T-cell receptor (TCR) signalling to exert their immunosuppressive activity, but the precise organization of the TCR signalling network compared to conventional T (Tconv) cells remains elusive. By using accurate mass spectrometry and multi-epitope ligand cartography (MELC) we characterized TCR signalling and recruitment of TCR signalling components to the immunological synapse (IS) in www.eji-journal.euThis is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. 2Marco van Ham et al. Eur. J. Immunol. 2017. 0: 1-16 in Treg cells, thereby, seems to be differentially organized as in Tconv cells: Treg cells show reduced Ca 2+ flux and ERK1/2 phosphorylation upon TCR stimulation [6][7][8], and downstream signalling molecules such as Lck, LAT and PLCγ1 are essential for Treg cell suppressive capacity [9,10]. In this line, a novel TCRmediated ADAP/integrin-independent PLCγ1 activation pathway was described to be required for suppression of Tconv cells by Treg cells [11]. Furthermore, Treg cells exhibit reduced S473 phosphorylation of Akt which seems a prerequisite for suppression [12]. Although the enzymatic activity of the tyrosine kinase ZAP70 seems to be dispensable for the suppressive phenotype of Foxp3 + Treg cells [13], a single mutation within the SH2-domain of ZAP70 leads to impaired suppressive activities, which indicated its importance at the immunological synapse (IS) [14]. Finally, it has only recently been recognized that the recruitment of TCR signalling components into the IS is differentially organized in Treg cells and Tconv cells. The protein kinase PKCθ is recruited to the IS in Tconv cells, in Treg cells, however, this kinase was found to be sequestered away from the IS, but still of importance to control suppression [15]. The spatial recruitment of signalling components during IS formation critically depends on cytoskeleton dynamics, which in turn is controlled by TCR activation and subsequent phosphorylation of proteins regulating cytoskeleton reorganization [5]. As part of these processes the microtubule-organizing centre (MTOC) is rapidly translocated in proximity to the IS. MTOC repositioning depends on LAT, ZAP70 and SLP76 [16] and is regulated by a cascade of distinct isoforms of the family of novel protein kinase C (nPKC) [17]. The MTOC serves as a platform to coordinate molecular movements from and to the IS through the support of microtubule (MT) motors [18]. For instance, TCR microclusters move along MTs towards the centre of the IS in a dynein-dependent manner [19], and hindrance of MTOC polarization, molecular transport and cytoskeleton dynamics prevent proper propagation of TCR signals [20].It is now tempting to speculate that the localization of signalling modules within the IS may be instrumental for the formation of a Treg cell-specific IS and the suppressive phenotype of Treg cells. At this moment, however,...

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Retinoic Acid-Induced Protein 14 (RAI14) Promotes mTOR-Mediated Inflammation Under Inflammatory Stress and Chemical Hypoxia in a U87 Glioblastoma Cell Line

Shen

Zhang

et al. 2018

Cell Mol Neurobiol

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

Cited by 11 publications

References 56 publications

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

Priming of transcriptional memory responses via the chromatin accessibility landscape in T cells

TCR signalling network organization at the immunological synapses of murine regulatory T cells

Retinoic Acid-Induced Protein 14 (RAI14) Promotes mTOR-Mediated Inflammation Under Inflammatory Stress and Chemical Hypoxia in a U87 Glioblastoma Cell Line

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