Transcription Factors: Methods and Protocols

ISBN-13: 9781607617372 / Angielski / Twarda / 2010 / 428 str.

Paul J. Higgins

Opis

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 can use 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 highlight protocols that focus on specific transcription factor family members with particular relevance to human disease. Composed in the highly successful Methods in Molecular BiologyT 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 text on transcriptional regulation today.

Spis treści

Table of Contents Preface Contributors Part I. Transcription Factor Trafficking and Post-Translational Modifications: Overviews of Mechanisms 1. A review of post-translational modifications and sub-cellular localization of Ets transcription factors: possible connection with cancer and involvement in the hypoxic reponse Celine Charlot, Helene Dubois-Pot, Tsvetan Serchov, Yves Tourette And Bohdan Wasylyk 2. Regulation of transcription factor function by targeted protein degradation: an overview focusing on p53, c-Myc and c-jun Jukka Westermarck 3. Review of molecular mechanisms involved in the activation of the Nrf2-ARE signaling pathway by chemopreventive agents Aldo Giudice, Claudio Arra and Maria C. Turco Part II. Cytoplasmic-nuclear trafficking of transcription factors 4. Subnuclear localization and intranuclear trafficking of transcription factors Sayyed K. Zaidi, Ricardo F. Medina, Shirwin M. Pockwinse, Rachit Bakshi, Krishna P. Kota, Syed A. Ali, Daniel W. Young, Jeffery A. Nickerson, Amjad Javed, Martin Montecino, Andre J. van Wijnen, Jane B. Lian, Janet L. Stein and Gary S. Stein 5. Analysis of ligand dependent nuclear accumulation of Smads in TGF-b signaling Douglas A. Chapnick and Xuedong Liu 6. Raf/MEK/MAPK signaling stimulates the nuclear translocation and trans- activating activity of FOXM1 Richard Y.M. Ma, Tommy H.K. Tong, Wai Ying Leung and Kwok-Ming Yao 7. Coupling of dephosphorylation and nuclear export of Smads in TGF-b signaling Fangyan Dai, Xueyan Duan, Yao-Yun Liang, Xia Lin and Xin-Hua Feng 8. Assessing sequence-specific DNA binding and transcriptional activity of STAT1 transcription factor Thomas Meyer and Uwe Vinkemeier 9. Analysis of nuclear export using photoactivatable GFP fusion proteins and interspecies heterokaryons Kerry-Ann Nakrieko, Iordanka A. Ivanova and Lina Dagnino 10. Determination of nuclear localization signal sequences for Kruppel-like factor 8 Tina S. Mehta, Farah Monzur and Jihe Zhao 11. Methods to measure nuclear export of beta-catenin using fixed and live cell assays Manisha Sharma and Beric R. Henderson 12. Imaging of transcription factor trafficking in living cells: lessons from corticosteroid receptor dynamics Mayumi Nishi Part III. Post-Translational Modifications and Impact on Function 13. Hypoxia-inducible factors: posttranslational crosstalk of signalling pathways Elitsa Y. Dimova and Thomas Kiezmann 14. The basic helix-loop-helix leucine-zipper gene Mitf: analysis of alternative promoter choice and splicing Kapil Bharti, Julien Debbache, Xin Wang and Heinz Arnheiter 15. Phosphorylation control of nuclear receptors Sebastien Lalevee, Christine Ferry and Cecile Rochette-Egly 16. Regulation of Krüpple-like factor 5 by targeted protein degradation Ceshi Chen. 17. Post-translational control of ETS transcription factors: detection of modified factors at target gene promoters Li Li, Janice Saxton and Pete E. Shaw 18. Integration of protein kinases into transcription complexes: identifying components of immobilized in vitro pre-initiation complexes Hong-Mei Zhang, Stephanie Vouier, Glenn Hodgson and Peter E, Shaw 19. Posttranslational modification of p53 by ubiquitin Chunhong Yan 20. Phosphorylation-dependent

Blurb

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 can use 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 highlight protocols 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 text on transcriptional regulation today.