Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Differential regulation of gene expression by two isoforms of the human transcriptional coactivator MKL1

ABSTRACT: We identified two isoforms of human MKL1 that differ in their N-terminal domains. Since MKL1 is a transcriptional coactivator of SRF and regulates many SRF target genes, we wanted to analyze if transcription is differentially regulated by the two isoforms upon stimulation of the Rho-actin-MKL1-SRF pathway. Activity of the Rho-actin-MKL1 pathway was induced in EcR293 cell lines stably over-expressing the empty vector control, a 5’UTR-full length MKL1_S construct, or a 5’UTR-full length MKL1_L construct by LPA treatment.

ORGANISM(S): Homo sapiens

SUBMITTER: Tim Roloff

PROVIDER: E-GEOD-47209 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

TGF-β-induced differentiation into myofibroblasts involves specific regulation of two MKL1 isoforms.

Scharenberg Matthias A MA Pippenger Benjamin E BE Sack Ragna R Zingg Dominik D Ferralli Jacqueline J Schenk Susanne S Martin Ivan I Chiquet-Ehrismann Ruth R

Journal of cell science 20140114 Pt 5

Cellular transformation into myofibroblasts is a central physiological process enabling tissue repair. Its deregulation promotes fibrosis and carcinogenesis. TGF-β is the main inducer of the contractile gene program that drives myofibroblast differentiation from various precursor cell types. Crucial regulators of this transcriptional program are serum response factor (SRF) and its cofactor MKL1 (also known as MRTF-A). However, the exact mechanism of the crosstalk between TGF-β signaling and MKL1 ...[more]

PMID: 24424023

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Project description:We have previously shown that HC11 mammary epithelial cells react strongly to the overexpression of megakaryoblastic leukemia-1 (Mkl1) with induction of tenascin-C expression. The present study was designed to find genes co-regulated with tenascin-C by Mkl1 in a SAP domain-dependent manner and without the involvement of serum response factor (SRF). For this purpose, we compared the transcriptomes of three stable HC11 cell strains that either overexpress full length Mkl1-RFP (HC11-FL), Mkl1-RFP with a mutated SRF-interaction site (HC11-mutB1) or Mkl1-RFP with a deletion of the SAP domain (HC11-ΔSAP). HC11 cell strains stably expressing either FL-, mutB1- or ∆SAP-Mkl1-RFP clones were grown in triplicates in 0.03% FCS/RPMI medium without EGF for 48 h before total RNA was extracted. RNA was converted into labeled cDNA and hybridized to Affymetrix GeneChip Mouse Gene 1.0 ST arrays. RMA normalized expression values were calculated with the affy package from the Bioconductor 2.4 release (1) and differentially expressed genes were identified using moderated t-statistics calculated with the empirical Bayes method as implemented in the Bioconductor limma package (2). To be considered as differentially expressed between HC11-mutB1 and HC11-FL as well as HC11-∆SAP and HC11-FL cell lines, genes had to pass the filters: adjusted P-value ≤ 0.01 (with Benjamin-Hochberg false discovery correction), a minimum absolute linear fold change difference of 2.0 and a minimum average expression value of 4.0 (log2). References: (1) Gentleman, R., Carey, V., Bates, D., Bolstad, B., Dettling, M., Dudoit, S., Ellis, B., Gautier, L., Ge, Y., Gentry, J., Hornik, K., Hothorn, T., Huber, W., Iacus, S., Irizarry, R., Leisch, F., Li, C., Maechler, M., Rossini, A., Sawitzki, G., Smith, C., Smyth, G., Tierney, L., Yang, J., Zhang, J. (2004) Bioconductor: open software development for computational biology and bioinformatics. Genome Biology 5, R80 (2) Smyth, G. K., Speed, T. (2003) Normalization of cDNA microarray data. Methods 31, 265-273

Dataset Information

Differential regulation of gene expression by two isoforms of the human transcriptional coactivator MKL1

Publications

TGF-β-induced differentiation into myofibroblasts involves specific regulation of two MKL1 isoforms.

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