Project description:Human myometrial cells taken from biopsies of pre-menopausal non-cancerous non-pregnant uteri were cultured in complete MEM D-valine medium plus 10% FCS, and stably-transfected with the pcDNA3.1/V5-His TOPO vector (Invitrogen) harbouring either CREB, CREM alpha, CREM tau2alpha, ATF2, ATF2-small gene or a control empty-vector. Total RNA was extracted using TRI-reagent (Sigma) and RNeasy columns (Qiagen), and 10 ug used in target preparation for array hybridization. Note that the data files for GSM17040 and GSM17041 are identical
Project description:Human myometrial cells taken from biopsies of pre-menopausal non-cancerous non-pregnant uteri were cultured in complete MEM D-valine medium plus 10% FCS, and stably-transfected with the pcDNA3.1/V5-His TOPO vector (Invitrogen) harbouring either CREB, CREM alpha, CREM tau2alpha, ATF2, ATF2-small gene or a control empty-vector. Total RNA was extracted using TRI-reagent (Sigma) and RNeasy columns (Qiagen), and 10 ug used in target preparation for array hybridization. Keywords: other
Project description:CREM (cAMP responsive element modulator) together with CREB and ATF-1 belong to the CREB family of transcriptional factors, that respond to cyclic AMP signaling and bind to cAMP responsive element (CRE) sites in promoters of selected genes. CREM can produce isoforms that have either activating or repressing functions, depending on the transcription of specific exons. In testis, it is involved in the regulation of spermatogenesis. In this dataset, we include the expression data obtained from wild-type and Crem knock-out mouse testis. 2 condition experiment: 2 strains (WT, Crem-/-). 5 biological replications per condition.
Project description:CREM (cAMP responsive element modulator) together with CREB and ATF-1 belong to the CREB family of transcriptional factors, that respond to cyclic AMP signaling and bind to cAMP responsive element (CRE) sites in promoters of selected genes. CREM can produce isoforms that have either activating or repressing functions, depending on the transcription of specific exons. In adrenal glands it is involved in the regulation of expression of genes for steroid hormone synthesis. In this dataset, we include the expression data obtained from wild-type and Crem knock-out mouse adrenal glands. 2 condition experiment: 2 strains (WT, Crem-/-). 5 biological replications per condition.
Project description:CREM (cAMP responsive element modulator) together with CREB and ATF-1 belong to the CREB family of transcriptional factors, that respond to cyclic AMP signaling and bind to cAMP responsive element (CRE) sites in promoters of selected genes. CREM can produce isoforms that have either activating or repressing functions, depending on the transcription of specific exons. In this dataset, we include the expression data obtained from wild-type (WT) and Crem knock-out (KO) mouse liver. Several comparisons were made. KO<WT at 0h, KO<WT at 12 h, 0h<12h in WT and 0h<12h in KO. 4 condition experiment: 2 strains (WT, Crem-/-), 2 time points (0h and 12h). 5 biological replications per condition.
Project description:Gene expression profiling of immortalized human mesenchymal stem cells with hTERT/E6/E7 transfected MSCs. hTERT may change gene expression in MSCs. Goal was to determine the gene expressions of immortalized MSCs.
Project description:Kynureninase is a member of a large family of catalytically diverse but structurally homologous pyridoxal 5'-phosphate (PLP) dependent enzymes known as the aspartate aminotransferase superfamily or alpha-family. The Homo sapiens and other eukaryotic constitutive kynureninases preferentially catalyze the hydrolytic cleavage of 3-hydroxy-l-kynurenine to produce 3-hydroxyanthranilate and l-alanine, while l-kynurenine is the substrate of many prokaryotic inducible kynureninases. The human enzyme was cloned with an N-terminal hexahistidine tag, expressed, and purified from a bacterial expression system using Ni metal ion affinity chromatography. Kinetic characterization of the recombinant enzyme reveals classic Michaelis-Menten behavior, with a Km of 28.3 +/- 1.9 microM and a specific activity of 1.75 micromol min-1 mg-1 for 3-hydroxy-dl-kynurenine. Crystals of recombinant kynureninase that diffracted to 2.0 A were obtained, and the atomic structure of the PLP-bound holoenzyme was determined by molecular replacement using the Pseudomonas fluorescens kynureninase structure (PDB entry 1qz9) as the phasing model. A structural superposition with the P. fluorescens kynureninase revealed that these two structures resemble the "open" and "closed" conformations of aspartate aminotransferase. The comparison illustrates the dynamic nature of these proteins' small domains and reveals a role for Arg-434 similar to its role in other AAT alpha-family members. Docking of 3-hydroxy-l-kynurenine into the human kynureninase active site suggests that Asn-333 and His-102 are involved in substrate binding and molecular discrimination between inducible and constitutive kynureninase substrates.
Project description:CREM (cAMP responsive element modulator) together with CREB and ATF-1 belong to the CREB family of transcriptional factors, that respond to cyclic AMP signaling and bind to cAMP responsive element (CRE) sites in promoters of selected genes. CREM can produce isoforms that have either activating or repressing functions, depending on the transcription of specific exons. In testis, it is involved in the regulation of spermatogenesis. In this dataset, we include the expression data obtained from wild-type and Crem knock-out mouse testis.
Project description:CREM (cAMP responsive element modulator) together with CREB and ATF-1 belong to the CREB family of transcriptional factors, that respond to cyclic AMP signaling and bind to cAMP responsive element (CRE) sites in promoters of selected genes. CREM can produce isoforms that have either activating or repressing functions, depending on the transcription of specific exons. In adrenal glands it is involved in the regulation of expression of genes for steroid hormone synthesis. In this dataset, we include the expression data obtained from wild-type and Crem knock-out mouse adrenal glands.
Project description:Ablation of the Creb1 gene in forebrain neurons was performed using the Cre/loxP system, with the recombinase expressed from the Camk2alfa promoter. Mice were crossed into the Crem KO background to prevent compensation of CREB loss by CREM overexpression. Our goal was to analyze how loss of CREB will affect acitivity-regulated transcription induced by strong stimulation, i.e. cocaine. Keywords: Treatment x Genotype