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Mammalian TAF(II)30 is required for cell cycle progression and specific cellular differentiation programmes
D Metzger1, E Scheer, A Soldatov
1Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, Collège de France, BP 163-67404 Illkirch Cedex, CU de Strasbourg, France.
Abstract:
The two alleles of the 30 kDa TATA-binding protein associated factor (TAF(II)30) gene, have been targeted by homologous recombination in murine F9 embryonal carcinoma cells and subsequently disrupted using a Cre recombinase-loxP strategy. The TAF(II)30-null cells are not viable, but are rescued by the expression of human TAF(II)30. Cells lacking TAF(II)30 are blocked in G(1)/G(0) phase of the cell cycle and undergo apoptosis. In agreement with the G(1) arrest phenotype, the expression of cyclin E is impaired and the retinoblastoma protein is hypophosphorylated in the TAF(II)30-null cells. Interestingly, retinoic acid (RA) treatment prevented TAF(II)30-null cell death and induced primitive endodermal differentiation. In contrast, the RA- and cAMP-induced parietal endodermal differentiation was impaired in the TAF(II)30-null cells. Thus, TAF(II)30 is not indispensable for class II gene transcription in general, but seems to be required for the expression of a subset of genes.
Insights
The TAF(II)30 gene is essential for cell viability, causing cell cycle arrest and apoptosis when disrupted. Retinoic acid rescues these cells, highlighting TAF(II)30
Area of Science:
- Molecular Biology
- Cell Biology
- Gene Regulation
Background:
- TATA-binding protein associated factors (TAFs) are crucial components of the TFIID complex, involved in transcription initiation.
- TAF(II)30 is a specific TAF implicated in regulating gene expression.
Purpose of the Study:
- To investigate the essentiality of TAF(II)30 in cell viability and cell cycle progression.
- To determine the role of TAF(II)30 in differentiation pathways.
Main Methods:
- Homologous recombination and Cre-loxP strategy were used to disrupt the TAF(II)30 gene in murine F9 embryonal carcinoma cells.
- Cell viability, cell cycle phase, apoptosis, protein phosphorylation, and gene expression were analyzed.
- Retinoic acid (RA) and cAMP treatments were employed to study differentiation.
Main Results:
- TAF(II)30-null cells exhibited G(1)/G(0) cell cycle arrest and apoptosis, but were rescued by human TAF(II)30 expression.
- Impaired cyclin E expression and hypophosphorylated retinoblastoma protein were observed in null cells.
- RA treatment prevented cell death and induced primitive endodermal differentiation in null cells.
- RA and cAMP-induced parietal endodermal differentiation was impaired in TAF(II)30-null cells.
Conclusions:
- TAF(II)30 is essential for the viability of F9 embryonal carcinoma cells.
- TAF(II)30 plays a critical role in regulating the G(1)/G(0) cell cycle transition and preventing apoptosis.
- TAF(II)30 is required for specific differentiation pathways, indicating its role in a subset of gene transcription.