In the last few years, significant breakthroughs in transcription research expanded our appreciation for the complexity of molecular controls on gene expression in mammalian cells. In Transcription Factors: Methods and Protocols , experts in the field describe state-of-the-art approaches that investigators canuse to probe critical mechanisms underlying transcription factor nuclear-cytoplasmic trafficking as well as to assess the functional impact of post-translational modifications on transcription factor function. The chapters are written by prominent scientists, many of whom developed these methods, and highlightprotocols that focus on specific transcription factor family members with particular relevance to human disease. Composed in the highly successful Methods in Molecular Biology™ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Comprehensive and current, Transcription Factors: Methods and Protocols compiles the latest techniques for elucidating controls on transcription factor intracellular localization and activity, and consequently is unlike any other methods-based texton transcriptional regulation today. The methods chapters are preceded by several excellent reviews that highlight the role of post-translational modifications and subcellular trafficking in transcription factor function Includes newtechniques for assessing kinase targeting of specific transcription factors, sites of phosphorylation and the involved signaling pathways Describes novel molecular and elegant microscopic imaging-based procedures to evaluate nuclear translocation and export in living cells INDICE: A review of post-translational modifications and sub-cellular localization of Ets transcription factors: possible connection with cancer and involvement in the hypoxic reponse.- Regulation of transcription factor function by targeted protein degradation: an overview focusing on p53, c-Myc and c-jun.- Review of molecular mechanisms involved in the activation of the Nrf2-ARE signaling pathway by chemopreventive agents.- Subnuclear localization and intranuclear trafficking of transcription factors.- Analysis of ligand dependent nuclear accumulation of Smads in TGF-b signaling.- Raf/MEK/MAPK signaling stimulates the nuclear translocation and trans- activating activity of FOXM.- Coupling of dephosphorylation and nuclear export of Smads in TGF-b signaling.- Assessing sequence-specific DNA binding and transcriptional activity of STAT1 transcription factor.- Analysis of nuclear export using photoactivatable GFP fusion proteins and interspecies heterokaryons.- Determination of nuclear localization signal sequences for Kruppel-like factor 8.- Methods to measure nuclear export of beta-catenin using fixed and live cell assays.- Imaging of transcriptionfactor trafficking in living cells: lessons from corticosteroid receptor dynamics.- Hypoxia-inducible factors: posttranslational crosstalk of signalling pathways.- The basic helix-loop-helix leucine-zipper gene Mitf: analysis of alternative promoter choice and splicing.- Phosphorylation control of nuclear receptors.- Regulation of Krüpple-like factor 5 by targeted protein degradation.- Post-translational control of ETS transcription factors: detection of modifiedfactors at target gene promoters.- Integration of protein kinases into transcription complexes: identifying components of immobilized in vitro pre-initiation complexes.- Posttranslational modification of p53 by ubiquitin.- Phosphorylation-dependent regulation of SATB1, the higher-order chromatin organizer and global gene regulator.- In vivo and in vitro tools to identify and study transcriptional regulation of USF-1 target genes.- Measuring the absolute abundanceof the Smad transcription factors using quantitative immunoblotting.- Flow cytometry analysis of transcription factors in T lymphocytes.- Identification ofspecific Protein/E-box-containing DNA complexes: lessons from the ubiquitously expressed USF transcription factors of the b-HLH-LZ super family.
- ISBN: 978-1-60761-737-2
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 377
- Fecha Publicación: 29/06/2010
- Nº Volúmenes: 1
- Idioma: Inglés