Analysis of the heat shock factor complex in mammalian HSP70 promoter.
Methods Mol Biol
; 1292: 53-65, 2015.
Article
em En
| MEDLINE
| ID: mdl-25804747
ABSTRACT
The heat shock response is characterized by the induction of heat shock proteins (HSPs) and is one of prominent mechanisms that regulate proteostasis capacity in the cell. In mammals, heat shock factor 1 (HSF1) regulates the expression of HSPs transcriptionally in both unstressed and stressed cells. Recent reports show that the HSF1-RPA complex constitutively gains access to nucleosomal DNA in part by recruiting a histone chaperone and a chromatin-remodeling component. Here, we describe the strategies to substitute endogenous HSF1 with ectopically expressed HSF1 or its mutant and to detect the occupancy of HSF1 transcription complex including RPA in vivo on two heat shock response elements located close together in the human or mouse HSP70 promoters by chromatin immunoprecipitation assay with high sensitivity and specificity.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Bioensaio
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Regiões Promotoras Genéticas
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Proteínas de Choque Térmico HSP70
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Complexos Multiproteicos
Limite:
Animals
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Humans
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article