Transcription of the caspase-14 gene in human epidermal keratinocytes requires AP-1 and NFκB

doi:10.1016/j.bbrc.2008.04.050

Biochemical and Biophysical Research Communications

Volume 371, Issue 2, 27 June 2008, Pages 261-266

https://doi.org/10.1016/j.bbrc.2008.04.050 Get rights and content

Abstract

Caspase-14, a protease involved in skin barrier formation, is specifically expressed in epidermal keratinocytes (KCs). Here, we mapped three start sites of transcription of the human caspase-14 gene and analyzed the upstream chromosomal region for promoter activity. Reporter gene assays identified a core promoter region proximal to the first exon and a distal regulatory region which differentially suppressed promoter activity in KC and other cells. Sequence elements in the proximal promoter were bound by the transcription factors AP-1 (JunB, c-Jun, JunD, Fra-1 and Fra-2) and NFκB (p50 and RelB). Our data reveal the basic organization of the human caspase-14 promoter and suggest an important role of AP-1 and NFκB in the transcriptional control of caspase-14.

Section snippets

Materials and methods

In silicopromoter analysis. Nucleotide sequences were obtained from the GenBank (human chromosome 19, build 36 version 2 of the NCBI’s genome annotation). The caspase-14 promoter region was investigated using the algorithms TFSEARCH (Parallel Application TRC Laboratory, RWCP, Japan) and IFTI-MIRAGE Tfsitescan.

Cell culture and in vitro organotypic skin cultures. KCs derived from neonatal foreskin of single donors were purchased from Lonza (Basel, Switzerland). The secondary human lung fibroblast

Cloning and promoter analysis of the 5′-end of the human caspase-14 gene

In a previous report, we have shown that the open reading frame of human caspase-14 is localized on six exons which are preceded on the 5′-side by an additional exon that lacks coding sequence [6]. Here, we have extended these studies by performing primer extension assays and RLM-RACE PCR on RNA from differentiated human primary KCs to map the start site(s) of transcription. Both methods yielded the same results and only RACE data are presented here. Transcription of caspase-14 was initiated at

Discussion

Our study defines the proximal promoter of human caspase-14 and implicates the two transcription factors AP-1 and NFκB in the basal transcriptional regulation of caspase-14.

AP-1 is a dimeric protein complex comprising members of the Jun and Fos proto-oncogene subfamilies in various combinations [23]. JunB, c-jun and, to a lower extent, JunD as well as fra-1 and fra-2 bound to the caspase-14 AP-1 site. The efficient suppression of caspase-14 expression by specific silencing of JunB indicated

Acknowledgments

We thank Wim Declercq (Ghent University, Belgium) for helpful discussions and Heidemarie Rossiter for critical reading of the manuscript.

References (28)

L. Eckhart et al.
Terminal differentiation of human keratinocytes and stratum corneum formation is associated with caspase-14 activation
J. Invest. Dermatol.
(2000)
L. Eckhart et al.
Caspase-14: analysis of gene structure and mRNA expression during keratinocyte differentiation
Biochem .Biophys. Res. Commun.
(2000)
H. Fischer et al.
Caspase-14 but not caspase-3 is processed during the development of fetal mouse epidermis
Differentiation
(2005)
S. Lippens et al.
Vitamin D3 induces caspase-14 expression in psoriatic lesions and enhances caspase-14 processing in organotypic skin cultures
Am. J. Pathol.
(2004)
M. Rendl et al.
Caspase-14 expression by epidermal keratinocytes is regulated by retinoids in a differentiation-associated manner
J. Invest. Dermatol.
(2002)
A. Geusau et al.
2,3,7,8-Tetrachlorodibenzo-p-dioxin impairs differentiation of normal human epidermal keratinocytes in a skin equivalent model
J. Invest. Dermatol.
(2005)
D.S. Walsh et al.
Psoriasis is characterized by altered epidermal expression of caspase 14, a novel regulator of keratinocyte terminal differentiation and barrier formation
J. Dermatol. Sci.
(2005)
M. Mildner et al.
Gene silencing in a human organotypic skin model
Biochem. Biophys. Res. Commun.
(2006)
H. Harant et al.
Synergistic activation of interleukin-8 gene transcription by all-trans-retinoic acid and tumor necrosis factor-alpha involves the transcription factor NF-kappaB
J. Biol. Chem.
(1996)
H. Fischer et al.
Stratum corneum-derived caspase-14 is catalytically active
FEBS Lett.
(2004)

J.F. Welter et al.

Fos-related antigen (Fra-1), junB, and junD activate human involucrin promoter transcription by binding to proximal and distal AP1 sites to mediate phorbol ester effects on promoter activity

J. Biol. Chem.

(1995)

A. Gazel et al.

Transcriptional profiling of epidermal keratinocytes: comparison of genes expressed in skin, cultured keratinocytes, and reconstituted epidermis, using large DNA microarrays

J. Invest. Dermatol.

(2003)

M. Karin et al.

AP-1 function and regulation

Curr. Opin. Cell Biol.

(1997)

S.I. Jang et al.

Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene

J. Biol. Chem.

(1996)

Cited by (13)

Biophysical insights into OR2T7: Investigation of a potential prognostic marker for glioblastoma
2022, Biophysical Journal
Citation Excerpt :
In addition, the expressions of c-Fos, c-Jun, and JunB have been extensively implicated in the progression of gliomas (60–64). p38 MAPK also targets Casp14 and RhoB transcription factors, which include c-Jun and JunB, to their promoter sites (65–69). Casp14 and RhoB have also been shown to suppress tumor growth (67,69–76), although in some cases they have been associated with promoting tumor growth (77–79).
Glioblastoma multiforme (GBM) is the most aggressive and prevalent form of brain cancer, with an expected survival of 12–15 months following diagnosis. GBM affects the glial cells of the central nervous system, which impairs regular brain function including memory, hearing, and vision. GBM has virtually no long-term survival even with treatment, requiring novel strategies to understand disease progression. Here, we identified a somatic mutation in OR2T7, a G-protein-coupled receptor (GPCR), that correlates with reduced progression-free survival for glioblastoma (log rank p-value = 0.05), suggesting a possible role in tumor progression. The mutation, D125V, occurred in 10% of 396 glioblastoma samples in The Cancer Genome Atlas, but not in any of the 2504 DNA sequences in the 1000 Genomes Project, suggesting that the mutation may have a deleterious functional effect. In addition, transcriptome analysis showed that the p38α mitogen-activated protein kinase (MAPK), c-Fos, c-Jun, and JunB proto-oncogenes, and putative tumor suppressors RhoB and caspase-14 were underexpressed in glioblastoma samples with the D125V mutation (false discovery rate < 0.05). Molecular modeling and molecular dynamics simulations have provided preliminary structural insight and indicate a dynamic helical movement network that is influenced by the membrane-embedded, cytofacial-facing residue 125, demonstrating a possible obstruction of G-protein binding on the cytofacial exposed region. We show that the mutation impacts the “open” GPCR conformation, potentially affecting G_α-subunit binding and associated downstream activity. Overall, our findings suggest that the Val125 mutation in OR2T7 could affect glioblastoma progression by downregulating GPCR-p38 MAPK tumor-suppression pathways and impacting the biophysical characteristics of the structure that facilitates G_α-subunit binding. This study provides the theoretical basis for further experimental investigation required to confirm that the D125V mutation in OR2T7 is not a passenger mutation. With validation, the aforementioned mutation could represent an important prognostic marker and a potential therapeutic target for glioblastoma.
An updated view on the functions of caspases in inflammation and immunity
2018, Seminars in Cell and Developmental Biology
Citation Excerpt :
Last, caspase-14 is restricted to the skin, both in humans and mice, where it functions in keratinocyte differentiation [217,218]. Whether it exerts roles in inflammation is not clear but its expression is induced in human psoriasis patients lesions [219] and is regulated by NF-κB and AP-1 [220]. Furthermore, Casp14−/− mice has a mild skin microbiota dysbiosis [221], which could stem from direct or indirect regulatory function of caspase-14 in mucosal immunity.
The binary classification of mammalian caspases as either apoptotic or inflammatory is now obsolete. Emerging data indicate that all mammalian caspases are intricately involved in the regulation of inflammation and immunity. They participate in embryonic and adult tissue homeostasis, control leukocyte differentiation, activation and effector functions, and mediate innate and adaptive immunity signaling. Caspases also promote host resistance by regulating anti-oxidant defense and pathogen clearance through regulation of phagosomal maturation, actin dynamics and phagosome-lysosome fusion. Beyond apoptosis, they regulate inflammatory cell death, eliciting rapid pyroptosis of infected cells, while inhibiting necroptosis-mediated tissue destruction and chronic inflammation. In this review, we describe the cellular and molecular mechanisms underlying non-apoptotic functions of caspases in inflammation and immunity and provide an updated view of their functions as central regulators of tissue homeostasis and host defense.
Caspases Connect Cell-Death Signaling to Organismal Homeostasis
2016, Immunity
Citation Excerpt :
Fragmentary evidence suggests that CASP14 may also participate in the regulation of inflammatory responses in the skin. Indeed, CASP14 is under the control of several transcription factors with pro- and anti-inflammatory functions; such transcription factors include NF-κB (which controls CASP14 expression in human epidermal keratinocytes) (Ballaun et al., 2008) and nuclear receptor subfamily 3, group C, member 1 (NR3C1, best known as glucocorticoid receptor, which influences Casp14 activation in the epidermis of late mouse embryos) (Bayo et al., 2008). Moreover, psoriasis (a skin disorder with a prominent inflammatory component) has been associated with altered expression of CASP14 in patient biopsies (Walsh et al., 2005), and the skin of Casp14−/− mice exhibits a mild form of dysbiosis as compared to that of wild-type mice (Kubica et al., 2014), possibly indicating an alteration in the immunological control of symbiotic bacteria.
Some forms of regulated cell death, such as apoptosis, are precipitated by the activation of cysteine proteases of the caspase family, including caspase 8, 9, and 3. Other caspases, such as caspase 1 and 4, are well known for their pro-inflammatory functions but regulate cell death in a limited number of pathophysiological settings. Accumulating evidence suggests that the most conserved function of mammalian caspases is not to control cell death sensu stricto, but to regulate inflammatory and immune reactions to dying cells and infectious challenges. Here, we review the molecular and cellular mechanisms though which mammalian caspases connect cell-death signaling to the maintenance of organismal homeostasis.
A guide for building biological pathways along with two case studies: Hair and breast development
2015, Methods
Citation Excerpt :
Casp14 is known to play a role in apoptosis, and in addition, it is directly associated with epidermal cell differentiation [76,77]. Casp14 is also upregulated by NF-kappaB and FOS, although downregulated by JUN [78]. However, both upregulators are suppressed by GPRC5D and NR3C1, respectively [79,80].
Genomic information is being underlined in the format of biological pathways. Building these biological pathways is an ongoing demand and benefits from methods for extracting information from biomedical literature with the aid of text-mining tools. Here we hopefully guide you in the attempt of building a customized pathway or chart representation of a system. Our manual is based on a group of software designed to look at biointeractions in a set of abstracts retrieved from PubMed. However, they aim to support the work of someone with biological background, who does not need to be an expert on the subject and will play the role of manual curator while designing the representation of the system, the pathway. We therefore illustrate with two challenging case studies: hair and breast development. They were chosen for focusing on recent acquisitions of human evolution. We produced sub-pathways for each study, representing different phases of development. Differently from most charts present in current databases, we present detailed descriptions, which will additionally guide PESCADOR users along the process. The implementation as a web interface makes PESCADOR a unique tool for guiding the user along the biointeractions, which will constitute a novel pathway.
ROS and NF-κB are involved in upregulation of IL-8 in A549 cells exposed to multi-walled carbon nanotubes
2009, Biochemical and Biophysical Research Communications
Citation Excerpt :
Western blotting and electrophoretic mobility shift assay (EMSA). Preparation of cell lysates and Western blotting were performed as described previously [19]. Nuclear extracts from A549 cells incubated with MWCNTs were prepared as described previously [20].
Carbon nanotubes (CNTs) have potential applications in biosensors, tissue engineering, and biomedical devices because of their unique physico-chemical, electronic and mechanical properties. However, there is limited literature data available concerning the biological properties and toxicity of CNTs. This study aimed to assess the toxicity exhibited by multi-walled CNTs (MWCNTs) and to elucidate possible molecular mechanisms underlying the biological effects of MWCNTs in A549 cells. Exposing A549 cells to MWCNTs led to cell death, changes in cell size and complexity, reactive oxygen species (ROS) production, interleukin-8 (IL-8) gene expression and nuclear factor (NF)-κB activation. Treatment of A549 cells with antioxidants prior to adding MWCNTs decreased ROS production and abrogated expression of IL-8 mRNA. Pretreatment of A549 cells with NF-κB inhibitors suppressed MWCNTs-induced IL-8 mRNA expression. These results indicate that MWCNTs are able to induce expression of IL-8 in A549 cells, at least in part, mediated by oxidative stress and NF-κB activation.
Pyroptosis: Role and Mechanisms in Cardiovascular Disease
2022, Frontiers in Cardiovascular Medicine

View all citing articles on Scopus

¹: Present address: Immunology Program, The Wistar Institute, Philadelphia, PA 19104, USA.

²: Present address: Children’s Cancer Research Institute, A-1090 Vienna, Austria.

View full text

Transcription of the caspase-14 gene in human epidermal keratinocytes requires AP-1 and NFκB

Abstract

Section snippets

Materials and methods

Cloning and promoter analysis of the 5′-end of the human caspase-14 gene

Discussion

Acknowledgments

J. Invest. Dermatol.

Biochem .Biophys. Res. Commun.

Differentiation

Am. J. Pathol.

J. Invest. Dermatol.

J. Invest. Dermatol.

J. Dermatol. Sci.

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

FEBS Lett.

J. Biol. Chem.

J. Invest. Dermatol.

Curr. Opin. Cell Biol.

J. Biol. Chem.