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FGF8, c-Abl and p300 participate in a pathway that controls stability and function of the ΔNp63α protein.

Restelli M, Molinari E, Marinari B, Conte D, Gnesutta N, Costanzo A, Merlo GR, Guerrini L - Hum. Mol. Genet. (2015)

Bottom Line: Here we show that the p300 acetyl-transferase physically interacts in vivo with ΔNp63α and catalyzes its acetylation on lysine 193 (K193) inducing ΔNp63α stabilization and activating specific transcriptional functions.Notably, the natural mutant ΔNp63α-K193E, associated to the Split-Hand/Foot Malformation-IV syndrome, cannot be acetylated by this pathway.Our results link FGF8, c-Abl and p300 in a regulatory pathway that controls ΔNp63α protein stability and transcriptional activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences, Università degli Studi di Milano, 20133 Milano, Italy.

No MeSH data available.


Related in: MedlinePlus

The ΔNp63α-K193E mutant displays an altered DNA binding activity and transcriptional activity on developmental related genes. (A) Expression of CASP10, EGFR, PERP and p53 was analyzed by Real-Time qPCR in U2OS cells stably transfected with pCDNA3 (empty vector), ΔNp63α or ΔNp63α-K193E cDNAs. (B) ΔNp63α and ΔNp63α-K193E proteins expression was confirmed by WB analysis. (C) Cells used in A and B were subjected to ChIP analysis, and the recovered chromatin was amplified with PERP, EGFR, p53 and CASP10-specific primers.
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DDV151F6: The ΔNp63α-K193E mutant displays an altered DNA binding activity and transcriptional activity on developmental related genes. (A) Expression of CASP10, EGFR, PERP and p53 was analyzed by Real-Time qPCR in U2OS cells stably transfected with pCDNA3 (empty vector), ΔNp63α or ΔNp63α-K193E cDNAs. (B) ΔNp63α and ΔNp63α-K193E proteins expression was confirmed by WB analysis. (C) Cells used in A and B were subjected to ChIP analysis, and the recovered chromatin was amplified with PERP, EGFR, p53 and CASP10-specific primers.

Mentions: To further characterize the transcriptional activity of the ΔNp63α-K193E mutant, we performed real-time, quantitative qPCR analyses in U2OS cells stably transfected with either the wild-type ΔNp63α or the ΔNp63α-K193E expression vectors. Interestingly, we confirmed that the ΔNp63α-K193E mutant over-expression results in altered expression of ΔNp63α target genes involved in development and apoptosis such as PERP, CASP10, EGFR (18,50) while it behaves like the wild-type ΔNp63α on p53 (Fig. 6A). Taken together, these data clearly show that the K193E mutation alters the transcriptional activity of ΔNp63α in a gene-specific manner.Figure 6.


FGF8, c-Abl and p300 participate in a pathway that controls stability and function of the ΔNp63α protein.

Restelli M, Molinari E, Marinari B, Conte D, Gnesutta N, Costanzo A, Merlo GR, Guerrini L - Hum. Mol. Genet. (2015)

The ΔNp63α-K193E mutant displays an altered DNA binding activity and transcriptional activity on developmental related genes. (A) Expression of CASP10, EGFR, PERP and p53 was analyzed by Real-Time qPCR in U2OS cells stably transfected with pCDNA3 (empty vector), ΔNp63α or ΔNp63α-K193E cDNAs. (B) ΔNp63α and ΔNp63α-K193E proteins expression was confirmed by WB analysis. (C) Cells used in A and B were subjected to ChIP analysis, and the recovered chromatin was amplified with PERP, EGFR, p53 and CASP10-specific primers.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492388&req=5

DDV151F6: The ΔNp63α-K193E mutant displays an altered DNA binding activity and transcriptional activity on developmental related genes. (A) Expression of CASP10, EGFR, PERP and p53 was analyzed by Real-Time qPCR in U2OS cells stably transfected with pCDNA3 (empty vector), ΔNp63α or ΔNp63α-K193E cDNAs. (B) ΔNp63α and ΔNp63α-K193E proteins expression was confirmed by WB analysis. (C) Cells used in A and B were subjected to ChIP analysis, and the recovered chromatin was amplified with PERP, EGFR, p53 and CASP10-specific primers.
Mentions: To further characterize the transcriptional activity of the ΔNp63α-K193E mutant, we performed real-time, quantitative qPCR analyses in U2OS cells stably transfected with either the wild-type ΔNp63α or the ΔNp63α-K193E expression vectors. Interestingly, we confirmed that the ΔNp63α-K193E mutant over-expression results in altered expression of ΔNp63α target genes involved in development and apoptosis such as PERP, CASP10, EGFR (18,50) while it behaves like the wild-type ΔNp63α on p53 (Fig. 6A). Taken together, these data clearly show that the K193E mutation alters the transcriptional activity of ΔNp63α in a gene-specific manner.Figure 6.

Bottom Line: Here we show that the p300 acetyl-transferase physically interacts in vivo with ΔNp63α and catalyzes its acetylation on lysine 193 (K193) inducing ΔNp63α stabilization and activating specific transcriptional functions.Notably, the natural mutant ΔNp63α-K193E, associated to the Split-Hand/Foot Malformation-IV syndrome, cannot be acetylated by this pathway.Our results link FGF8, c-Abl and p300 in a regulatory pathway that controls ΔNp63α protein stability and transcriptional activity.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosciences, Università degli Studi di Milano, 20133 Milano, Italy.

No MeSH data available.


Related in: MedlinePlus