Abstract
Investigation of DNA-protein interactions is a key approach in understanding mechanisms of gene regulation. The method described allows detection of dynamic DNA-protein interactions occurring at gene promoters in living cells during the time scale of seconds and minutes. The combination of chromatin immunoprecipitation with real-time PCR allows for detection of changes in activator and co-activator content of any promoter during transcriptional activation. The described method is most applicable to investigation of processes resulting in nucleosome loss at gene promoters during the induction of transcription. The approach is also applicable to any dynamic process involving DNA-protein interactions.
Publication types
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Chromatin Assembly and Disassembly / genetics
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Chromatin Assembly and Disassembly / physiology*
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Chromatin Immunoprecipitation / methods*
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Heat-Shock Proteins / genetics
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Heat-Shock Proteins / metabolism
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Histones / metabolism
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Protein Binding
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RNA Polymerase II / genetics
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RNA Polymerase II / metabolism
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Real-Time Polymerase Chain Reaction / methods*
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / metabolism
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Transcriptional Activation
Substances
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Heat-Shock Proteins
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Histones
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Saccharomyces cerevisiae Proteins
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RNA Polymerase II