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Dynamics of the full length and mutated heat shock factor 1 in human cells.
- Source :
-
PloS one [PLoS One] 2013 Jul 08; Vol. 8 (7), pp. e67566. Date of Electronic Publication: 2013 Jul 08 (Print Publication: 2013). - Publication Year :
- 2013
-
Abstract
- Heat shock factor 1 is the key transcription factor of the heat shock response. Its function is to protect the cell against the deleterious effects of stress. Upon stress, HSF1 binds to and transcribes hsp genes and repeated satellite III (sat III) sequences present at the 9q12 locus. HSF1 binding to pericentric sat III sequences forms structures known as nuclear stress bodies (nSBs). nSBs represent a natural amplification of RNA pol II dependent transcription sites. Dynamics of HSF1 and of deletion mutants were studied in living cells using multi-confocal Fluorescence Correlation Spectroscopy (mFCS) and Fluorescence Recovery After Photobleaching (FRAP). In this paper, we show that HSF1 dynamics modifications upon heat shock result from both formation of high molecular weight complexes and increased HSF1 interactions with chromatin. These interactions involve both DNA binding with Heat Shock Element (HSE) and sat III sequences and a more transient sequence-independent binding likely corresponding to a search for more specific targets. We find that the trimerization domain is required for low affinity interactions with chromatin while the DNA binding domain is required for site-specific interactions of HSF1 with DNA.
- Subjects :
- Animals
Cell Membrane Permeability
Cell Nucleus Structures metabolism
Chemical Fractionation
DNA metabolism
DNA-Binding Proteins chemistry
Diffusion
Fluorescence Recovery After Photobleaching
Gene Knockdown Techniques
Green Fluorescent Proteins metabolism
HSP70 Heat-Shock Proteins metabolism
HeLa Cells
Heat Shock Transcription Factors
Heat-Shock Response genetics
Humans
Intracellular Space metabolism
Mice
Molecular Weight
Mutant Proteins chemistry
Protein Binding
Protein Structure, Tertiary
Protein Transport
RNA metabolism
Spectrometry, Fluorescence
Stress, Physiological
Subcellular Fractions metabolism
Transcription Factors chemistry
Transcriptional Activation genetics
DNA-Binding Proteins genetics
DNA-Binding Proteins metabolism
Mutant Proteins metabolism
Mutation genetics
Transcription Factors genetics
Transcription Factors metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 8
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 23861773
- Full Text :
- https://doi.org/10.1371/journal.pone.0067566