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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 expression in NHEK cells (A) NHEK cells were induced to differentiate and RNA was used to perform RTqPCR. Scotin doubles its expression (from 0 to 5 days) in differentiating human keratinocytes. (B) TAp63α is upregulated at the transcriptional level while (C) ΔNp63α is down-regulated as expected in differentiating NHEK cells. (D) Western blot experiments performed on human keratinocyte lysates showed Scotin induction at day 3 and 5 of differentiation. This is paralleled by ΔNp63 down-regulation (Ab4 clone antibody) and keratin 10 upregulation. (E) The densitometric analysis, that is representative of few experiments, shows that Scotin expression is increased by almost 50% in differentiating NHEK cells.
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fig4: Scotin expression in NHEK cells (A) NHEK cells were induced to differentiate and RNA was used to perform RTqPCR. Scotin doubles its expression (from 0 to 5 days) in differentiating human keratinocytes. (B) TAp63α is upregulated at the transcriptional level while (C) ΔNp63α is down-regulated as expected in differentiating NHEK cells. (D) Western blot experiments performed on human keratinocyte lysates showed Scotin induction at day 3 and 5 of differentiation. This is paralleled by ΔNp63 down-regulation (Ab4 clone antibody) and keratin 10 upregulation. (E) The densitometric analysis, that is representative of few experiments, shows that Scotin expression is increased by almost 50% in differentiating NHEK cells.

Mentions: In order to confirm our data in man, we tested Scotin expression in a primary epithelial cell line (NHEK) induced to differentiate in vitro; RTqPCR analysis showed that Scotin expression doubles by day 5 during differentiation (Fig. 4A), and this phenomenon is clearly paralleled by increased expression of TAp63α (Fig. 4B) with a reduction in ΔNp63α expression level (Fig. 4C). Western blot analysis performed on keratinocyte lysates shows that there is almost a fifty per cent induction of Scotin at the protein level during epidermal differentiation (Fig. 4D and E). Again K10 demonstrates that NHEK cells fully undergo the differentiation process.


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 expression in NHEK cells (A) NHEK cells were induced to differentiate and RNA was used to perform RTqPCR. Scotin doubles its expression (from 0 to 5 days) in differentiating human keratinocytes. (B) TAp63α is upregulated at the transcriptional level while (C) ΔNp63α is down-regulated as expected in differentiating NHEK cells. (D) Western blot experiments performed on human keratinocyte lysates showed Scotin induction at day 3 and 5 of differentiation. This is paralleled by ΔNp63 down-regulation (Ab4 clone antibody) and keratin 10 upregulation. (E) The densitometric analysis, that is representative of few experiments, shows that Scotin expression is increased by almost 50% in differentiating NHEK cells.
© Copyright Policy
Related In: Results  -  Collection

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

fig4: Scotin expression in NHEK cells (A) NHEK cells were induced to differentiate and RNA was used to perform RTqPCR. Scotin doubles its expression (from 0 to 5 days) in differentiating human keratinocytes. (B) TAp63α is upregulated at the transcriptional level while (C) ΔNp63α is down-regulated as expected in differentiating NHEK cells. (D) Western blot experiments performed on human keratinocyte lysates showed Scotin induction at day 3 and 5 of differentiation. This is paralleled by ΔNp63 down-regulation (Ab4 clone antibody) and keratin 10 upregulation. (E) The densitometric analysis, that is representative of few experiments, shows that Scotin expression is increased by almost 50% in differentiating NHEK cells.
Mentions: In order to confirm our data in man, we tested Scotin expression in a primary epithelial cell line (NHEK) induced to differentiate in vitro; RTqPCR analysis showed that Scotin expression doubles by day 5 during differentiation (Fig. 4A), and this phenomenon is clearly paralleled by increased expression of TAp63α (Fig. 4B) with a reduction in ΔNp63α expression level (Fig. 4C). Western blot analysis performed on keratinocyte lysates shows that there is almost a fifty per cent induction of Scotin at the protein level during epidermal differentiation (Fig. 4D and E). Again K10 demonstrates that NHEK cells fully undergo the differentiation process.

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