Abstract
Signal transducer and activator of transcription 3 (Stat3) is a key regulator of gene expression in response to signaling of the glycoprotein 130 (gp130) family cytokines, including interleukin 6, oncostatin M, and leukemia inhibitory factor. Many efforts have been made to identify Stat3 target genes and to understand the mechanism of how Stat3 regulates gene expression. Using the microarray technique, hundreds of genes have been documented to be potential Stat3 target genes in different cell types. However, only a small fraction of these genes have been proven to be true direct Stat3 target genes. Here we report the identification of novel direct Stat3 target genes using a genome-wide screening procedure based on the chromatin immunoprecipitation method. These novel Stat3 target genes are involved in a diverse array of biological processes such as oncogenesis, cell growth, and differentiation. We show that Stat3 can act as both a repressor and activator on its direct target genes. We further show that most of the novel Stat3 direct target genes are dependent on Stat3 for their transcriptional regulation. In addition, using a physiological cell system, we demonstrate that Stat3 is required for the transcriptional regulation of two of the newly identified direct Stat3 target genes important for muscle differentiation.
Highlights
NIH3T3 cells were treated with oncostatin M (OSM) for 30 min, and chromatin immunoprecipitation (ChIP) assays were performed with a Signal transducer and activator of transcription 3 (Stat3)-specific antibody
The precipitated genomic DNA fragments from the ChIP were amplified by ligation-mediated PCR and further subcloned into pBluescript followed by sequencing of the cloned inserts
Using the ChIP method with an antibody specific for Stat3 followed by ligation-mediated PCR and subcloning, we have identified a whole new set of Stat3 target genes involved in a wide ranging array of grown in growth media and transfected with Stat3 siRNA or cellular processes
Summary
LIF-mediated self-renewal of murine embryonic stem cells requires Stat activation [21,22,23] Because of these wide ranging physiological functions of Stat, it is critical to identify Stat target genes to fully understand how Stat mediates its effect on these various cellular processes. It is essential to identify direct Stat target genes to develop specific therapeutic treatments as well as to understand how Stat regulates gene transcription to achieve diverse physiological impacts. A significant number of novel Stat target genes were identified by this method These novel genes are diverse in their function, including involvement in oncogenesis, neuronal development, and muscle differentiation. We have identified a set of novel direct Stat target genes important for a wide range of biological processes, including the control of cellular growth and differentiation
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