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Regulation of the expression and processing of caspase-12.

Kalai M, Lamkanfi M, Denecker G, Boogmans M, Lippens S, Meeus A, Declercq W, Vandenabeele P - J. Cell Biol. (2003)

Bottom Line: The effect is increased further when IFN-gamma is combined with TNF, lipopolysaccharide (LPS), or dsRNA.Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis.Similar processing occurs in TNF-, LPS-, Fas ligand-, and thapsigargin (Tg)-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biomedical Research, Unit of Molecular Signalling and Cell Death, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium.

ABSTRACT
Phylogenetic analysis clusters caspase-12 with the inflammatory caspases 1 and 11. We analyzed the expression of caspase-12 in mouse embryos, adult organs, and different cell types and tested the effect of interferons (IFNs) and other proinflammatory stimuli. Constitutive expression of the caspase-12 protein was restricted to certain cell types, such as epithelial cells, primary fibroblasts, and L929 fibrosarcoma cells. In fibroblasts and B16/B16 melanoma cells, caspase-12 expression is stimulated by IFN-gamma but not by IFN-alpha or -beta. The effect is increased further when IFN-gamma is combined with TNF, lipopolysaccharide (LPS), or dsRNA. These stimuli also induce caspase-1 and -11 but inhibit the expression of caspase-3 and -9. In contrast to caspase-1 and -11, no caspase-12 protein was detected in macrophages in any of these treatments. Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis. Similar processing occurs in TNF-, LPS-, Fas ligand-, and thapsigargin (Tg)-induced apoptosis. However, B16/B16 melanoma cells die when treated with the ER stress-inducing agent Tg whether they express caspase-12 or not.

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Immunohistochemical analysis of the expression of caspase-12 in the mouse embryo. Caspase-12 expression was detected in sections using either anti–caspase-12 rabbit antiserum Ab-1 or Ab-2. Results obtained with both antisera are similar (not depicted). Only those obtained with Ab-2 are presented. Negative controls of adjacent sections were stained by the anti–caspase-12 antisera depleted with recombinant caspase-12. Sections stained by secondary antibody alone were completely negative (not depicted). (A) Sections of a 13.5-d-postcoitum embryo. Note the strong staining of heart, lung, and nose. White box shows the embryo at its relative real size (6 mm). (B) A close-up of the heart (1) in A. (C) A close-up of the lung (2) in A. Note the positive staining of the epithelial layer of the trachea. (D) A close-up of the nose (3) in A. Note the positive staining of the epithelium in the entrance of the nose. Bars: (A) 455 μm; (B) 80 μm; (C) 30 μm; (D) 45 μm.
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fig3: Immunohistochemical analysis of the expression of caspase-12 in the mouse embryo. Caspase-12 expression was detected in sections using either anti–caspase-12 rabbit antiserum Ab-1 or Ab-2. Results obtained with both antisera are similar (not depicted). Only those obtained with Ab-2 are presented. Negative controls of adjacent sections were stained by the anti–caspase-12 antisera depleted with recombinant caspase-12. Sections stained by secondary antibody alone were completely negative (not depicted). (A) Sections of a 13.5-d-postcoitum embryo. Note the strong staining of heart, lung, and nose. White box shows the embryo at its relative real size (6 mm). (B) A close-up of the heart (1) in A. (C) A close-up of the lung (2) in A. Note the positive staining of the epithelial layer of the trachea. (D) A close-up of the nose (3) in A. Note the positive staining of the epithelium in the entrance of the nose. Bars: (A) 455 μm; (B) 80 μm; (C) 30 μm; (D) 45 μm.

Mentions: Next, we analyzed, by Western blotting, extracts from seven murine cell lines representing different cell types (B cell hybridoma, T cells, preB cells, fibrosarcoma, melanoma, and promyelocytes). A control lysate of HEK293T cells transiently transfected with a plasmid leading to overexpression of caspase-12 revealed the 49-kD band expected for full-length pro–caspase-12 and two additional bands running at 38- and 28-kD, respectively (Fig. 2 B). Caspase-12 protein was detected only in cells derived from the murine fibrosarcoma cell line L929 (Fig. 2 B). In agreement with the apparent absence of caspase-12 in lymphoid organs, no caspase-12 protein was detected in cell lines of hematopoietic origin. The strongest expression was detected in L929r2, the cells from which the cDNA of caspase-12 was originally cloned (Van de Craen et al., 1997). Analysis of 13.5-d-postcoitum mouse embryos by immunohistochemistry revealed that the most positively stained organ is the heart (Fig. 3, A and B). In several other organs, including the lung and the nose, the expression of the protein was confined to certain cell types or layers, such as epithelium of the trachea and the nose trail (Fig. 3, C and D). These results suggest that the constitutive expression of caspase-12 is probably restricted to certain cell types.


Regulation of the expression and processing of caspase-12.

Kalai M, Lamkanfi M, Denecker G, Boogmans M, Lippens S, Meeus A, Declercq W, Vandenabeele P - J. Cell Biol. (2003)

Immunohistochemical analysis of the expression of caspase-12 in the mouse embryo. Caspase-12 expression was detected in sections using either anti–caspase-12 rabbit antiserum Ab-1 or Ab-2. Results obtained with both antisera are similar (not depicted). Only those obtained with Ab-2 are presented. Negative controls of adjacent sections were stained by the anti–caspase-12 antisera depleted with recombinant caspase-12. Sections stained by secondary antibody alone were completely negative (not depicted). (A) Sections of a 13.5-d-postcoitum embryo. Note the strong staining of heart, lung, and nose. White box shows the embryo at its relative real size (6 mm). (B) A close-up of the heart (1) in A. (C) A close-up of the lung (2) in A. Note the positive staining of the epithelial layer of the trachea. (D) A close-up of the nose (3) in A. Note the positive staining of the epithelium in the entrance of the nose. Bars: (A) 455 μm; (B) 80 μm; (C) 30 μm; (D) 45 μm.
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Related In: Results  -  Collection

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fig3: Immunohistochemical analysis of the expression of caspase-12 in the mouse embryo. Caspase-12 expression was detected in sections using either anti–caspase-12 rabbit antiserum Ab-1 or Ab-2. Results obtained with both antisera are similar (not depicted). Only those obtained with Ab-2 are presented. Negative controls of adjacent sections were stained by the anti–caspase-12 antisera depleted with recombinant caspase-12. Sections stained by secondary antibody alone were completely negative (not depicted). (A) Sections of a 13.5-d-postcoitum embryo. Note the strong staining of heart, lung, and nose. White box shows the embryo at its relative real size (6 mm). (B) A close-up of the heart (1) in A. (C) A close-up of the lung (2) in A. Note the positive staining of the epithelial layer of the trachea. (D) A close-up of the nose (3) in A. Note the positive staining of the epithelium in the entrance of the nose. Bars: (A) 455 μm; (B) 80 μm; (C) 30 μm; (D) 45 μm.
Mentions: Next, we analyzed, by Western blotting, extracts from seven murine cell lines representing different cell types (B cell hybridoma, T cells, preB cells, fibrosarcoma, melanoma, and promyelocytes). A control lysate of HEK293T cells transiently transfected with a plasmid leading to overexpression of caspase-12 revealed the 49-kD band expected for full-length pro–caspase-12 and two additional bands running at 38- and 28-kD, respectively (Fig. 2 B). Caspase-12 protein was detected only in cells derived from the murine fibrosarcoma cell line L929 (Fig. 2 B). In agreement with the apparent absence of caspase-12 in lymphoid organs, no caspase-12 protein was detected in cell lines of hematopoietic origin. The strongest expression was detected in L929r2, the cells from which the cDNA of caspase-12 was originally cloned (Van de Craen et al., 1997). Analysis of 13.5-d-postcoitum mouse embryos by immunohistochemistry revealed that the most positively stained organ is the heart (Fig. 3, A and B). In several other organs, including the lung and the nose, the expression of the protein was confined to certain cell types or layers, such as epithelium of the trachea and the nose trail (Fig. 3, C and D). These results suggest that the constitutive expression of caspase-12 is probably restricted to certain cell types.

Bottom Line: The effect is increased further when IFN-gamma is combined with TNF, lipopolysaccharide (LPS), or dsRNA.Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis.Similar processing occurs in TNF-, LPS-, Fas ligand-, and thapsigargin (Tg)-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Biomedical Research, Unit of Molecular Signalling and Cell Death, Ghent University, Ledeganckstraat 35, B-9000 Ghent, Belgium.

ABSTRACT
Phylogenetic analysis clusters caspase-12 with the inflammatory caspases 1 and 11. We analyzed the expression of caspase-12 in mouse embryos, adult organs, and different cell types and tested the effect of interferons (IFNs) and other proinflammatory stimuli. Constitutive expression of the caspase-12 protein was restricted to certain cell types, such as epithelial cells, primary fibroblasts, and L929 fibrosarcoma cells. In fibroblasts and B16/B16 melanoma cells, caspase-12 expression is stimulated by IFN-gamma but not by IFN-alpha or -beta. The effect is increased further when IFN-gamma is combined with TNF, lipopolysaccharide (LPS), or dsRNA. These stimuli also induce caspase-1 and -11 but inhibit the expression of caspase-3 and -9. In contrast to caspase-1 and -11, no caspase-12 protein was detected in macrophages in any of these treatments. Transient overexpression of full-length caspase-12 leads to proteolytic processing of the enzyme and apoptosis. Similar processing occurs in TNF-, LPS-, Fas ligand-, and thapsigargin (Tg)-induced apoptosis. However, B16/B16 melanoma cells die when treated with the ER stress-inducing agent Tg whether they express caspase-12 or not.

Show MeSH
Related in: MedlinePlus