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Genetics

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Iowa State University

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Full-Text Articles in Molecular Biology

Polytene Chromosome Squash Methods For Studying Transcription And Epigenetic Chromatin Modification In Drosophila Using Antibodies, Kristen M. Johansen, Weili Cai, Huai Deng, Xiaomin Bao, Weiguo Zhang, Jack Girton, Jorgen Johansen Aug 2009

Polytene Chromosome Squash Methods For Studying Transcription And Epigenetic Chromatin Modification In Drosophila Using Antibodies, Kristen M. Johansen, Weili Cai, Huai Deng, Xiaomin Bao, Weiguo Zhang, Jack Girton, Jorgen Johansen

Biochemistry, Biophysics and Molecular Biology Publications

The giant polytene chromosomes from Drosophila third instar larval salivary glands provide an important model system for studying the architectural changes in chromatin morphology associated with the process of transcription initiation and elongation. Especially, analysis of the heat shock response has proved useful in correlating chromatin structure remodeling with transcriptional activity. An important tool for such studies is the labeling of polytene chromosome squash preparations with antibodies to the enzymes, transcription factors, or histone modifications of interest. However, in any immunohistochemical experiment there will be advantages and disadvantages to different methods of fixation and sample preparation, the relative merits of ...


Rna Polymerase Ii-Mediated Transcription At Active Loci Does Not Require Histone H3s10 Phosphorylation In Drosophila, Weili Cai, Xiaomin Bao, Huai Deng, Ye Jin, Jack Girton, Jorgen Johansen, Kristen M. Johansen Jan 2008

Rna Polymerase Ii-Mediated Transcription At Active Loci Does Not Require Histone H3s10 Phosphorylation In Drosophila, Weili Cai, Xiaomin Bao, Huai Deng, Ye Jin, Jack Girton, Jorgen Johansen, Kristen M. Johansen

Biochemistry, Biophysics and Molecular Biology Publications

JIL-1 is the major kinase controlling the phosphorylation state of histone H3S10 at interphase in Drosophila. In this study, we used three different commercially available histone H3S10 phosphorylation antibodies, as well as an acid-free polytene chromosome squash protocol that preserves the antigenicity of the histone H3S10 phospho-epitope, to examine the role of histone H3S10 phosphorylation in transcription under both heat shock and non-heat shock conditions. We show that there is no redistribution or upregulation of JIL-1 or histone H3S10 phosphorylation at transcriptionally active puffs in such polytene squash preparations after heat shock treatment. Furthermore, we provide evidence that heat shock-induced ...