Large scale RNAi screen reveals that the inhibitor of DNA binding 2 (ID2) protein is repressed by p53 family member p63 and functions in human keratinocyte differentiation

Ning Wu, David Castel, Marie-Anne Debily, Maria Alessandra Vigano, Olivier Alibert, Roberto Mantovani, Kristiina Iljin, Paul-Henri Romeo, Xavier Gidrol (Corresponding Author)

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Abstract

The inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, ID2, acts as an oncogene and elevated levels of ID2 have been reported in several malignancies. Whereas some inducers of the ID2 gene have been characterized, little is known regarding the proteins capable to repress its expression. We developed siRNA microarrays to perform a large scale loss-of-function screen in human adult keratinocytes engineered to express GFP under the control of the upstream region of ID2 gene. We screened the effect of siRNA-dependent inhibition of 220 cancer-associated genes on the expression of the ID2::GFP reporter construct. Three genes NBN, RAD21, and p63 lead to a repression of ID2 promoter activity. Strikingly NBN and RAD21 are playing on major role in cell cycle progression and mitosis arrest. These results underline the pregnant need to silence ID2 expression at transcript level to promote cell cycle exit. Central to this inhibitory mechanism we find p63, a key transcription factor in epithelial development and differentiation, which binds specific cis-acting sequence within the ID2 gene promoter both in vitro and in vivo. P63 would not suppress ID2 expression, but would rather prevent excessive expression of that protein to enable the onset of keratinocyte differentiation.
Original languageEnglish
Pages (from-to)20870-20879
Number of pages10
JournalJournal of Biological Chemistry
Volume286
Issue number23
DOIs
Publication statusPublished - 2011
MoE publication typeA1 Journal article-refereed

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Inhibitor of Differentiation Protein 2
RNA Interference
Keratinocytes
DNA
Genes
Small Interfering RNA
Cell Cycle
Cells
Neoplasm Genes
Microarrays
Oncogenes
Mitosis

Cite this

Wu, Ning ; Castel, David ; Debily, Marie-Anne ; Vigano, Maria Alessandra ; Alibert, Olivier ; Mantovani, Roberto ; Iljin, Kristiina ; Romeo, Paul-Henri ; Gidrol, Xavier. / Large scale RNAi screen reveals that the inhibitor of DNA binding 2 (ID2) protein is repressed by p53 family member p63 and functions in human keratinocyte differentiation. In: Journal of Biological Chemistry. 2011 ; Vol. 286, No. 23. pp. 20870-20879.
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title = "Large scale RNAi screen reveals that the inhibitor of DNA binding 2 (ID2) protein is repressed by p53 family member p63 and functions in human keratinocyte differentiation",
abstract = "The inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, ID2, acts as an oncogene and elevated levels of ID2 have been reported in several malignancies. Whereas some inducers of the ID2 gene have been characterized, little is known regarding the proteins capable to repress its expression. We developed siRNA microarrays to perform a large scale loss-of-function screen in human adult keratinocytes engineered to express GFP under the control of the upstream region of ID2 gene. We screened the effect of siRNA-dependent inhibition of 220 cancer-associated genes on the expression of the ID2::GFP reporter construct. Three genes NBN, RAD21, and p63 lead to a repression of ID2 promoter activity. Strikingly NBN and RAD21 are playing on major role in cell cycle progression and mitosis arrest. These results underline the pregnant need to silence ID2 expression at transcript level to promote cell cycle exit. Central to this inhibitory mechanism we find p63, a key transcription factor in epithelial development and differentiation, which binds specific cis-acting sequence within the ID2 gene promoter both in vitro and in vivo. P63 would not suppress ID2 expression, but would rather prevent excessive expression of that protein to enable the onset of keratinocyte differentiation.",
author = "Ning Wu and David Castel and Marie-Anne Debily and Vigano, {Maria Alessandra} and Olivier Alibert and Roberto Mantovani and Kristiina Iljin and Paul-Henri Romeo and Xavier Gidrol",
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Large scale RNAi screen reveals that the inhibitor of DNA binding 2 (ID2) protein is repressed by p53 family member p63 and functions in human keratinocyte differentiation. / Wu, Ning; Castel, David; Debily, Marie-Anne; Vigano, Maria Alessandra; Alibert, Olivier; Mantovani, Roberto; Iljin, Kristiina; Romeo, Paul-Henri; Gidrol, Xavier (Corresponding Author).

In: Journal of Biological Chemistry, Vol. 286, No. 23, 2011, p. 20870-20879.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Large scale RNAi screen reveals that the inhibitor of DNA binding 2 (ID2) protein is repressed by p53 family member p63 and functions in human keratinocyte differentiation

AU - Wu, Ning

AU - Castel, David

AU - Debily, Marie-Anne

AU - Vigano, Maria Alessandra

AU - Alibert, Olivier

AU - Mantovani, Roberto

AU - Iljin, Kristiina

AU - Romeo, Paul-Henri

AU - Gidrol, Xavier

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AB - The inhibitor of DNA binding 2, dominant negative helix-loop-helix protein, ID2, acts as an oncogene and elevated levels of ID2 have been reported in several malignancies. Whereas some inducers of the ID2 gene have been characterized, little is known regarding the proteins capable to repress its expression. We developed siRNA microarrays to perform a large scale loss-of-function screen in human adult keratinocytes engineered to express GFP under the control of the upstream region of ID2 gene. We screened the effect of siRNA-dependent inhibition of 220 cancer-associated genes on the expression of the ID2::GFP reporter construct. Three genes NBN, RAD21, and p63 lead to a repression of ID2 promoter activity. Strikingly NBN and RAD21 are playing on major role in cell cycle progression and mitosis arrest. These results underline the pregnant need to silence ID2 expression at transcript level to promote cell cycle exit. Central to this inhibitory mechanism we find p63, a key transcription factor in epithelial development and differentiation, which binds specific cis-acting sequence within the ID2 gene promoter both in vitro and in vivo. P63 would not suppress ID2 expression, but would rather prevent excessive expression of that protein to enable the onset of keratinocyte differentiation.

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