Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Effect on gene expression upon deletion of Ada3

ABSTRACT: Ada3 (alteration/deficiency in activation) is a transcriptional adaptor that forms a core structural component of multiple HAT complexes. In order to gain insights into physiological roles of Ada3, we made a conditional knockout mouse for Ada3 which was early embryonic lethal. Deletion of Ada3 in MEFs by using Adenovirus-Cre showed changes in global histone acetylation. We performed microarray analysis after deletion of Ada3 in these cells to compare gene expression changes caused due to Ada3 loss Exponentially growing Ada3FL/FL MEFs were infected with control or Cre adenovirus. After 3 days of infection, we collected RNA from these cells to perform microarray analysis. The experiment was done using biological replicates

ORGANISM(S): Mus musculus

SUBMITTER: Vimla Band

PROVIDER: E-GEOD-37542 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Mammalian alteration/deficiency in activation 3 (Ada3) is essential for embryonic development and cell cycle progression.

Mohibi Shakur S Gurumurthy Channabasavaiah Basavaraju CB Nag Alo A Wang Jun J Mirza Sameer S Mian Yousaf Y Quinn Meghan M Katafiasz Bryan B Eudy James J Pandey Sanjit S Guda Chittibabu C Naramura Mayumi M Band Hamid H Band Vimla V

The Journal of biological chemistry 20120626 35

Ada3 protein is an essential component of histone acetyl transferase containing coactivator complexes conserved from yeast to human. We show here that germline deletion of Ada3 in mouse is embryonic lethal, and adenovirus-Cre mediated conditional deletion of Ada3 in Ada3(FL/FL) mouse embryonic fibroblasts leads to a severe proliferation defect which was rescued by ectopic expression of human Ada3. A delay in G(1) to S phase of cell cycle was also seen that was due to accumulation of Cdk inhibito ...[more]

PMID: 22736770

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Project description:DNA polymerase zeta (Polz) plays a key role in translesion DNA synthesis. Inactivation of the gene encoding its catalytic subunit (Rev3L) leads to embryonic lethality and abrogates cell proliferation in primary fibroblasts highlighting an essential role of Polz in mammalian cells. However, its biological function remains poorly understood. Here, we examined if the absence of Rev3L may impact on transcriptional program by performing microarray-based transcriptome profiling from Rev3L+/+ and Rev3L-/- MEFs. For this study, we used mouse embryonic fibroblasts (MEF) that are derived from mice harboring one allele with essential exons flanked by loxP sites, and either one inactivated allele of Rev3L (Rev3L−/lox) or, one wild type allele (Rev3L+/lox). Upon introduction of an adenovirus expressing Cre, floxed copy of Rev3L was deleted thus generating Rev3L−/Δ cells, and cell lines were SV40 Tag-immortalized (Lange et al., NAR 2012). We then used Rev3L−/Δ and Rev3L+/lox MEFs (named Rev3L-/- and Rev3L+/+ MEFs, respectively in this study). From two independent cell culture (A and B), total RNAs were extracted from Rev3L+/+ and Rev3L-/- MEFs and processed to determine the transcriptional program in these cell lines. For this study, we used mouse embryonic fibroblasts (MEF) that are derived from mice harboring one allele with essential exons flanked by loxP sites, and either one inactivated allele of Rev3L (Rev3L−/lox) or, one wild type allele (Rev3L+/lox). Upon introduction of an adenovirus expressing Cre, floxed copy of Rev3L was deleted thus generating Rev3L−/Δ cells, and cell lines were SV40 Tag-immortalized (Lange et al., NAR 2012). We then used Rev3L−/Δ and Rev3L+/lox MEFs (named Rev3L-/- and Rev3L+/+ MEFs, respectively in this study). From two independent cell culture (A and B), total RNAs were extracted from Rev3L+/+ and Rev3L-/- MEFs and processed to determine the transcriptional program in these cell lines with Agilent Mouse Genomic array (8x60k) with two replicates in dual color.

Dataset Information

Effect on gene expression upon deletion of Ada3

Publications

Mammalian alteration/deficiency in activation 3 (Ada3) is essential for embryonic development and cell cycle progression.

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