Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Krüppel-like factor 3 KO and rescue expression data in murine embyronic fibroblasts

ABSTRACT: Krüppel-like factor 3 (KLF3) is a transcriptional repressor that has roles in adipogenesis, B-cell maturation and erythropoiesis (for review see Pearson et al., 2012). We profiled gene expression in murine embryonic fibroblasts (MEFs) where Klf3 had been knocked-out and where the same cell had been rescued with Klf3 using microarrays. Klf3 KO murine embryonic fibroblast cell lines were produced from Klf3 KO mice. These cells were rescued with epitope tagged Klf3 (Klf3-V5) using retroviral delivery (murine stem cell virus). Stable clones were selected uner puromycin seletion and total RNA was taken for array

ORGANISM(S): Mus musculus

SUBMITTER: Jon Burdach

PROVIDER: E-GEOD-44744 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Regions outside the DNA-binding domain are critical for proper in vivo specificity of an archetypal zinc finger transcription factor.

Burdach Jon J Funnell Alister P W AP Mak Ka Sin KS Artuz Crisbel M CM Wienert Beeke B Lim Wooi F WF Tan Lit Yeen LY Pearson Richard C M RC Crossley Merlin M

Nucleic acids research 20131007 1

Transcription factors (TFs) are often regarded as being composed of a DNA-binding domain (DBD) and a functional domain. The two domains are considered separable and autonomous, with the DBD directing the factor to its target genes and the functional domain imparting transcriptional regulation. We examined an archetypal zinc finger (ZF) TF, Krüppel-like factor 3 with an N-terminal domain that binds the corepressor CtBP and a DBD composed of three ZFs at its C-terminus. We established a system to ...[more]

PMID: 24106088

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Project description:Visceral fat (VF) and subcutaneous fat (SF) are developmentally different tissues with different gene expression. Islet-1 (ISL1), a LIM-homeobox transcription factor with important developmental and regulatory function in islet, neural, and cardiac tissue, is virtually absent in SF but substantially expressed in the stromovascular [preadipocyte containing] fraction of VF; expression correlates negatively with adiposity in rodents and man. ISL1 expression is transiently increased in 3T3-L1 preadipocytes during early differentiation, suggesting a functional role. To examine the role of ISL1 in adipogenesis, we tested whether retroviral overexpression of ISL1 in 3T3-L1 preadipocytes affected their ability to differentiate into mature adipocytes. Terminal differentiation was assessed by Oil Red O [lipid droplet] staining and by immunoblot detection of adipocyte marker proteins, including aP2 and GLUT4. ISL1 significantly inhibited lipid droplet formation, reduced lipid accumulation (about 80% inhibition, p<0.05), and substantially inhibited aP2 and GLUT4 expression. ISL1 did not inhibit expression of C/EBPb and C/EBPd after induction of differentiation, but reduced PPARg and C/EBPa by >50% at both mRNA and protein level. In addition, the PPARg agonist, rosiglitazone, substantially rescued ISL1 inhibited adipogenesis in the absence of exogenous PPARg, and fully rescued in the presence of exogenous PPARg. In summary, ISL1 overexpression inhibited fat droplet formation, lipid accumulation, and adipocyte-specific gene expression; there was accompanying inhibition of C/EBPa, PPARg and downstream gene expression. We conclude that ISL1 overexpression inhibited adipocyte differentiation by inhibition of PPARg regulated gene expression. As abdominal obesity strongly correlates with insulin resistance, and cardiovascular risk, ISL1 up-regulation may impact abdominal obesity and its concomitant metabolic derangements. Total cellular RNA was isolated from 3T3-L1 cells expressing Flag-ISL1 or not at 48 h following treatment with differentiation cocktail. Individual RNA from biological triplicates was used for microarray analysis.

Dataset Information

Krüppel-like factor 3 KO and rescue expression data in murine embyronic fibroblasts

Publications

Regions outside the DNA-binding domain are critical for proper in vivo specificity of an archetypal zinc finger transcription factor.

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