Unknown,Transcriptomics,Genomics,Proteomics

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Global analysis of gene expression by SV40 T antigen in mouse embryo fibroblasts

ABSTRACT: SV40 large T antigen (TAg) contributes to cell transformation, in part, by targeting two well characterized tumor suppressors, pRb and p53. TAg expression affects the transcriptional circuits controlled by Rb and by p53. We have performed a microarray analysis to examine the global change in gene expression induced by wild-type TAg and TAg-mutants, in an effort to link changes in gene expression to specific transforming functions. For this analysis we have used MEFs expressing TAg or infected by SV40. Our analysis indicates that TAg can induce interferon-stimulated genes in MEFs and that this induction depends upon the LXCXE motif and p53 binding. Two independent wild-type MEFs and three independent MEF cell lines expressing TAg and the mutants were used. Additionally, three independent mock and SV40 infections were conduted in MEFs. Total RNA was isolated from with the RNeasy kit. Total RNA was processed by the Genomics and Proteomics Core Laboratories at the University of Pittsburgh and hybridized to the Affymetrix GeneChip Mouse Genome 430 2.0 array. MAS 5.0 was used to obtain the present/absent calls and CEL files were normalized by RMA to obtain log2 expression values.

ORGANISM(S): Mus musculus

SUBMITTER: Paul Cantalupo

PROVIDER: E-GEOD-20620 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Induction of interferon-stimulated genes by Simian virus 40 T antigens.

Rathi Abhilasha V AV Cantalupo Paul G PG Sarkar Saumendra N SN Pipas James M JM

Virology 20100807 2

Simian virus 40 (SV40) large T antigen (TAg) is a multifunctional oncoprotein essential for productive viral infection and for cellular transformation. We have used microarray analysis to examine the global changes in cellular gene expression induced by wild-type T antigen (TAg(wt)) and TAg-mutants in mouse embryo fibroblasts (MEFs). The expression profile of approximately 800 cellular genes was altered by TAg(wt) and a truncated TAg (TAg(N136)), including many genes that influence cell cycle, D ...[more]

PMID: 20692676

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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

Global analysis of gene expression by SV40 T antigen in mouse embryo fibroblasts

Publications

Induction of interferon-stimulated genes by Simian virus 40 T antigens.

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