Endogenous TBK1-mediated GABARAP-L2 and LC3C phosphorylation site identification
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ABSTRACT: In order to identify TBK1-mediated GABARAP-L2 and LC3C phosphorylation sites, cells were treated with TBK1 or control siRNA and TBK1 kinase activity was induced by treatment with CCCP.
Project description:TBK1 phosphorylates GABARAP-L2 S87/88. In order to test if this phosphomimetic mutation in GABARAP-L2 changes its binding to other proteins a mass spectrometry based interactome study was performed.
Project description:The purpose of the dataset is to analyze expression of genes induced by KRAS and regulated by TBK1; The proto-oncogene KRAS is mutated in a wide array of human cancers, most of which are aggressive and respond poorly to standard therapies. Although the identification of specific oncogenes has led to the development of clinically effective, molecularly targeted therapies in some cases, KRAS has remained refractory to this approach. An alternative strategy for targeting KRAS is to identify gene products that, when suppressed or inhibited, result in cell death only in the presence of an oncogenic allele. Here we have used systematic RNA interference (RNAi) to detect synthetic lethal partners of oncogenic KRAS and found that the non-canonical IkB kinase, TBK1, was selectively essential in cells that harbor mutant KRAS. Suppression of TBK1 induced apoptosis specifically in human cancer cell lines that depend on oncogenic KRAS expression. In these cells, TBK1 activated NF- B anti-apoptotic signals involving cREL and BCL-XL that were essential for survival, providing mechanistic insights into this synthetic lethal interaction. These observations identify TBK1 as a potential therapeutic target in KRAS mutant tumors and establish a general approach for the rational identification of co-dependent pathways in cancer. Experiment Overall Design: Knock out of TBK1 in the contect of KRAS activation (mutant) and control (WT)
Project description:The proto-oncogene KRAS is mutated in a wide array of human cancers, most of which are aggressive and respond poorly to standard therapies. Although the identification of specific oncogenes has led to the development of clinically effective, molecularly targeted therapies in some cases, KRAS has remained refractory to this approach. An alternative strategy for targeting KRAS is to identify gene products that, when suppressed or inhibited, result in cell death only in the presence of an oncogenic allele. Here we have used systematic RNA interference (RNAi) to detect synthetic lethal partners of oncogenic KRAS and found that the non-canonical IkB kinase, TBK1, was selectively essential in cells that harbor mutant KRAS. Suppression of TBK1 induced apoptosis specifically in human cancer cell lines that depend on oncogenic KRAS expression. In these cells, TBK1 activated NF- B anti-apoptotic signals involving cREL and BCL-XL that were essential for survival, providing mechanistic insights into this synthetic lethal interaction. These observations identify TBK1 as a potential therapeutic target in KRAS mutant tumors and establish a general approach for the rational identification of co-dependent pathways in cancer. This SuperSeries is composed of the following subset Series:; GSE17643: Profiling of immortalized human lung epithelial cells following oncogenic KRAS expression and TBK1 suppression; GSE17671: Profiling of immortalized human lung epithelial cells following infection with oncogenic KRAS (G12V) Experiment Overall Design: Refer to individual Series
Project description:This study was designed to identify endoglin (ENG)-dependent BMP2-responsive genes in the mouse fibroblastic cell line PDL-L2. PDL-L2 cells were treated with an siRNA for endoglin (siENG) or a control siRNA (siCont). After 48hr, the cells were exposed to recombinant human BMP-2 (250 ng/ml) or vehicle and cultured for 12 hr.
Project description:The nematode Auanema rhodensis is trioecious (co-occurrence of males, females and self-fertile hermaphrodites). To better understand its sex determination system, we have compared the transcriptomic profiles of early (L2) females, hermaphrodites and converted females (hermaphrodite-fated larvae induced to develop as females). Additionally, we sequenced the transcriptome of adult males and individuals from various stages and sexes (mixed stages samples) to compare global gene expression profiles along the assembled draft chromosomes of A. rhodensis (BioProject PRJEB29492). The RNA-seq data was also used to predict genes in the assembled genome. We generated three biological replicates for each RNA-seq condition (L2 females, L2 converted females, L2 hermaphrodites, males and mixed stages). Comparisons of the expression profiles of the L2 conditions was performed to identify genes potentially involved in the sexual differentiation process, using a standard RNA-seq comparison approach. Briefly, the cleaned reads were aligned to the genome using STAR, the abundance and identification of differentially expressed genes were assessed using FeatureCounts and DEseq2 (using as thresholds an absolute log2(Fold Change) >= 2 and an FDR <0.01).
Project description:Arsenic is a carcinogen that is known to induce cell transformation and tumor formation. Although studies have been performed to examine the modulation of signaling molecules caused by arsenic exposure, the molecular mechanisms by which arsenic causes cancer are still unclear. We hypothesized that arsenic alters gene expression leading to carcinogenesis in the lung. In this study, we examined global gene expression in response to 0.75 M-BM-5M arsenic treatment for 1-7 days in a rat lung epithelial cell line (L2) using an in-house 10k rat DNA microarray. One hundred thirty one genes were identified using the one-class statistical analysis of microarray (SAM) test. Of them, 33 genes had a fold change of M-bM-^IM-% 2 at least between two time points. These genes were then clustered into 5 groups using K-means cluster analysis based on their expression patterns. Seven selected genes, all associated with cancer, were confirmed by real-time PCR. These genes have functions directly or indirectly related to metabolism, glycolysis, cell proliferation and differentiation, and regulation of transcription, all of which may be involved in neoplastic transformation of cells. Our findings provide important insight for the future studies of arsenic-mediated lung cancer. Keywords: Lung, aresenic, L2, micorarray L2 cells were exposed with 0.75 uM of arsenite for 0, 1, 3, 5, and 7 days (C, D1, D3, D5 and D7). The samples were arranged for hybridization using a loop design. For each paired sample, dye flip and three biological replications were performed for each sample.