Project description:HT1080 fibrosarcoma cells were treated with cytostatic drugs and analysed on Affymetrix microarray to identify genes and pathways regulated by these compounds. The array images were processed using Affymetrix MAS 5.0 software. and normalized by global scaling or scaled to a polyA+ spike mask Keywords = Cytostatic drugs Keywords = Actinomycin D Keywords = Doxorubicin Keywords = Vincristine Keywords = apoptosis Keywords: ordered
Project description:HT1080 fibrosarcoma cells were treated with cytostatic drugs and analysed on Affymetrix microarray to identify genes and pathways regulated by these compounds. The array images were processed using Affymetrix MAS 5.0 software. and normalized by global scaling or scaled to a polyA+ spike mask
Project description:Investigation of whole genome gene expression level changes in HT1080 fibrosarcoma cell line after transfection CRABP1 gene and R131A CRABP1 mutant (arginine-alanine substitution in a protein active site, protein lacks the ability to interact with retinoic acid), compared to HT1080 line transfected with empty pLXSN vector. A twelve samples on one chip study using total RNA recovered from four separate cultures of HT1080 human fibrosarcoma cell line transfected with empty pLXSN vector, four HT1080 cell line cultures transfected with CRABP1 gene and four HT1080 cell line cultures transfected with R131A CRABP1 mutant. Each sample on a chip measures the expression level of 44,049 genes from human genome with three-fold technical redundancy.
Project description:We compare histone modifications, chromatin accessibility, and replication timing domain genome-wide in HCT116 colon cancer cells with its genetic derivative DKO cells which lack DNMT3B and DNMT1 activity and the fibrosarcoma cell line HT1080.
Project description:Investigation of whole genome gene expression level changes in HT1080 fibrosarcoma cell line after transfection CRABP1 gene and R131A CRABP1 mutant (arginine-alanine substitution in a protein active site, protein lacks the ability to interact with retinoic acid), compared to HT1080 line transfected with empty pLXSN vector.
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:As the evolution of miRNA genes has been found to be one of the important factors in formation of the modern type of man, we performed a comparative analysis of the evolution of miRNA genes in two archaic hominines, Homo sapiens neanderthalensis and Homo sapiens denisova, and elucidated the expression of their target mRNAs in bain.A comparative analysis of the genomes of primates, including species in the genus Homo, identified a group of miRNA genes having fixed substitutions with important implications for the evolution of Homo sapiens neanderthalensis and Homo sapiens denisova. The mRNAs targeted by miRNAs with mutations specific for Homo sapiens denisova exhibited enhanced expression during postnatal brain development in modern humans. By contrast, the expression of mRNAs targeted by miRNAs bearing variations specific for Homo sapiens neanderthalensis was shown to be enhanced in prenatal brain development.Our results highlight the importance of changes in miRNA gene sequences in the course of Homo sapiens denisova and Homo sapiens neanderthalensis evolution. The genetic alterations of miRNAs regulating the spatiotemporal expression of multiple genes in the prenatal and postnatal brain may contribute to the progressive evolution of brain function, which is consistent with the observations of fine technical and typological properties of tools and decorative items reported from archaeological Denisovan sites. The data also suggest that differential spatial-temporal regulation of gene products promoted by the subspecies-specific mutations in the miRNA genes might have occurred in the brains of Homo sapiens denisova and Homo sapiens neanderthalensis, potentially contributing to the cultural differences between these two archaic hominines.
Project description:The conserved TFIIH helicases XPB and XPD play key roles in transcription initiation and DNA repair. To investigate the functions of these helicases on a genome-wide scale, we performed ChIP-seq of endogenous XPB and XPD in the HT1080 human fibrosarcoma cell line.