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Scotin: A new p63 target gene expressed during epidermal differentiation.

Zocchi L, Bourdon JC, Codispoti A, Knight R, Lane DP, Melino G, Terrinoni A - Biochem. Biophys. Res. Commun. (2007)

Bottom Line: We detected Scotin upregulation in primary keratinocyte cell lines committed to differentiate.In this paper we also show that Scotin is expressed in the supra basal layer of the epidermis in parallel with TAp63, but not DeltaNp63 expression.We conclude that Scotin is a new p63 target gene induced during epithelial differentiation, a complex process that also involves ER stress induction.

View Article: PubMed Central - PubMed

Affiliation: IDI-IRCCS Biochemistry Laboratory, c/o University of Tor Vergata, Department of Experimental Medicine, Via Montpellier 1, 00133 Rome, Italy.

ABSTRACT
p63, a member of the p53 family, is transcribed from two different promoters giving rise to two different proteins: TAp63 that contains the N-terminal transactivation domain and DeltaN that lacks this domain. In this article we describe a new target gene Scotin induced by TAp63 during epithelial differentiation. This gene was previously isolated as a p53-inducible proapoptotic gene and the protein is located in the endoplasmic reticulum and in the nuclear membrane. Scotin expression is induced in response to endoplasmic reticulum (ER) stress in a p53 dependent or independent manner. We detected Scotin upregulation in primary keratinocyte cell lines committed to differentiate. In this paper we also show that Scotin is expressed in the supra basal layer of the epidermis in parallel with TAp63, but not DeltaNp63 expression. We conclude that Scotin is a new p63 target gene induced during epithelial differentiation, a complex process that also involves ER stress induction.

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Scotin and GADD153 expression in mouse skin sections (A) Scotin is expressed only in the suprabasal layer of the epidermis (red), but it is not expressed in the basal layer where ΔNp63 (Ab4 clone) is present (green). (B) Western blot experiments performed on mouse keratinocyte lysates induced to differentiate (0, 1, 3, and 5 days) revealed Scotin upregulation at the protein level. Conversely ΔNp63 expression is progressively reduced compared to tubulin expression, during the differentiation program. (C) Involucrin increases, as expected, its expression in differentiating mouse keratinocytes (from 0 to 5 days). Western blot of tubulin was used as a loading control. Bar = 20 μm.
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fig3: Scotin and GADD153 expression in mouse skin sections (A) Scotin is expressed only in the suprabasal layer of the epidermis (red), but it is not expressed in the basal layer where ΔNp63 (Ab4 clone) is present (green). (B) Western blot experiments performed on mouse keratinocyte lysates induced to differentiate (0, 1, 3, and 5 days) revealed Scotin upregulation at the protein level. Conversely ΔNp63 expression is progressively reduced compared to tubulin expression, during the differentiation program. (C) Involucrin increases, as expected, its expression in differentiating mouse keratinocytes (from 0 to 5 days). Western blot of tubulin was used as a loading control. Bar = 20 μm.

Mentions: To further investigate the importance of Scotin during epidermal differentiation we analyzed its expression in the mouse. To facilitate the analysis of Scotin expression, the skin of new born mice has been used, since it is hairless although the epidermis is fully differentiated. The results demonstrate that Scotin is expressed in the upper layers of the epidermis (where TAp63 is present), but is completely absent from the basal layer, where ΔNp63α is highly expressed (Fig. 3A). We used a pan-p63 antibody and ΔNp63α is almost 99% of total p63 expression in the skin; therefore TAp63α expression is not detectable in the upper layers of epidermis with this antibody. Western blots performed on mouse keratinocytes committed to differentiate, as demonstrated by involucrin positivity (Fig. 3C), revealed that the strong upregulation of Scotin during differentiation (Fig. 3B) is accompanied by a reduction in ΔNp63α expression level.


Scotin: A new p63 target gene expressed during epidermal differentiation.

Zocchi L, Bourdon JC, Codispoti A, Knight R, Lane DP, Melino G, Terrinoni A - Biochem. Biophys. Res. Commun. (2007)

Scotin and GADD153 expression in mouse skin sections (A) Scotin is expressed only in the suprabasal layer of the epidermis (red), but it is not expressed in the basal layer where ΔNp63 (Ab4 clone) is present (green). (B) Western blot experiments performed on mouse keratinocyte lysates induced to differentiate (0, 1, 3, and 5 days) revealed Scotin upregulation at the protein level. Conversely ΔNp63 expression is progressively reduced compared to tubulin expression, during the differentiation program. (C) Involucrin increases, as expected, its expression in differentiating mouse keratinocytes (from 0 to 5 days). Western blot of tubulin was used as a loading control. Bar = 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2691585&req=5

fig3: Scotin and GADD153 expression in mouse skin sections (A) Scotin is expressed only in the suprabasal layer of the epidermis (red), but it is not expressed in the basal layer where ΔNp63 (Ab4 clone) is present (green). (B) Western blot experiments performed on mouse keratinocyte lysates induced to differentiate (0, 1, 3, and 5 days) revealed Scotin upregulation at the protein level. Conversely ΔNp63 expression is progressively reduced compared to tubulin expression, during the differentiation program. (C) Involucrin increases, as expected, its expression in differentiating mouse keratinocytes (from 0 to 5 days). Western blot of tubulin was used as a loading control. Bar = 20 μm.
Mentions: To further investigate the importance of Scotin during epidermal differentiation we analyzed its expression in the mouse. To facilitate the analysis of Scotin expression, the skin of new born mice has been used, since it is hairless although the epidermis is fully differentiated. The results demonstrate that Scotin is expressed in the upper layers of the epidermis (where TAp63 is present), but is completely absent from the basal layer, where ΔNp63α is highly expressed (Fig. 3A). We used a pan-p63 antibody and ΔNp63α is almost 99% of total p63 expression in the skin; therefore TAp63α expression is not detectable in the upper layers of epidermis with this antibody. Western blots performed on mouse keratinocytes committed to differentiate, as demonstrated by involucrin positivity (Fig. 3C), revealed that the strong upregulation of Scotin during differentiation (Fig. 3B) is accompanied by a reduction in ΔNp63α expression level.

Bottom Line: We detected Scotin upregulation in primary keratinocyte cell lines committed to differentiate.In this paper we also show that Scotin is expressed in the supra basal layer of the epidermis in parallel with TAp63, but not DeltaNp63 expression.We conclude that Scotin is a new p63 target gene induced during epithelial differentiation, a complex process that also involves ER stress induction.

View Article: PubMed Central - PubMed

Affiliation: IDI-IRCCS Biochemistry Laboratory, c/o University of Tor Vergata, Department of Experimental Medicine, Via Montpellier 1, 00133 Rome, Italy.

ABSTRACT
p63, a member of the p53 family, is transcribed from two different promoters giving rise to two different proteins: TAp63 that contains the N-terminal transactivation domain and DeltaN that lacks this domain. In this article we describe a new target gene Scotin induced by TAp63 during epithelial differentiation. This gene was previously isolated as a p53-inducible proapoptotic gene and the protein is located in the endoplasmic reticulum and in the nuclear membrane. Scotin expression is induced in response to endoplasmic reticulum (ER) stress in a p53 dependent or independent manner. We detected Scotin upregulation in primary keratinocyte cell lines committed to differentiate. In this paper we also show that Scotin is expressed in the supra basal layer of the epidermis in parallel with TAp63, but not DeltaNp63 expression. We conclude that Scotin is a new p63 target gene induced during epithelial differentiation, a complex process that also involves ER stress induction.

Show MeSH