Refined Solution Structure and Dynamics of the DNA-binding Domain of the Heat Shock Factor fromKluyveromyces lactis

Damberger, Fred F.; Pelton, Jeffrey G.; Liu, C.; Cho, H.; Harrison, Celia J.; Nelson, Hillary C. M.; Wemmer, David E.

doi:10.1006/jmbi.1995.0649

Cited by 20 publications

(5 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The wild-type and wingless DNAbinding domains had similar spectral features in the α-helix/ β-sheet region of 200-220 nm (Fig. 3A), indicating that the proteins were well-folded and contained mixed αβ secondary structure characteristics, consistent with previous structural studies of the HSF DNA-binding domain (8)(9)(10). The wingless and wild-type proteins do differ slightly in the 230 nm range (Fig.…”

Section: Removal Of the Wing Motif Does Not Affect Structure Of The D...supporting

confidence: 86%

See 1 more Smart Citation

The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity

Cicero

Hubl²,

Harrison³

et al. 2001

Nucleic Acids Research

View full text Add to dashboard Cite

The yeast heat shock transcription factor (HSF) belongs to the winged helix family of proteins. HSF binds DNA as a trimer, and additional trimers can bind DNA co-operatively. Unlike other winged helix-turn-helix proteins, HSF's wing does not appear to contact DNA, as based on a previously solved crystal structure. Instead, the structure implies that the wing is involved in protein-protein interactions, possibly within a trimer or between adjacent trimers. To understand the function of the wing in the HSF DNA-binding domain, a Saccharomyces cerevisiae strain was created that expresses a wingless HSF protein. This strain grows normally at 30 degrees C, but shows a decrease in reporter gene expression during constitutive and heat-shocked conditions. Removal of the wing does not affect the stability or trimeric nature of a protein fragment containing the DNA-binding and trimerization domains. Removal of the wing does result in a decrease in DNA-binding affinity. This defect was mainly observed in the ability to form the first trimer-bound complex, as the formation of larger complexes is unaffected by the deletion. Our results suggest that the wing is not involved in the highly co-operative nature of HSF binding, but may be important in stabilizing the first trimer bound to DNA.

show abstract

Section: Removal Of the Wing Motif Does Not Affect Structure Of The D...supporting

confidence: 86%

“…Crystallographic and NMR studies have shown that most residues of the yeast HSF wing were dynamic and presumably flexible in both the apo and complex structures (7)(8)(9)(10). However, one model predicted that the wing provides an interface for protein-protein contacts when bound to DNA (7).…”

Section: Introductionmentioning

confidence: 99%

The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity

Cicero

Hubl²,

Harrison³

et al. 2001

Nucleic Acids Research

View full text Add to dashboard Cite

show abstract

“…This kink is quite important, as seen in Figure 4g. It is present in the two hsf domains considered here, as well as in another structure of Kluyveromyves lactis hsf (Damberger et al, 1995). It seems to be due to the presence of a Pro residue near the helix C terminus.…”

Section: Group Related To the Homeodomain Family (Figures 2 3f)mentioning

confidence: 86%

Structural Classification of HTH DNA-binding Domains and Protein – DNA Interaction Modes

Wintjens

Rooman

1996

Journal of Molecular Biology

234

209

View full text Add to dashboard Cite

“…Although it is formally possible that the L189P mutation alters the overall structure of HSF1 in such a fashion that it affects the structure and function of the DNA binding domain, our findings that this mutant is fully functional in yeast, binds to the IL-6 promoter similar to the wild-type protein and is still able to provide some level of protection from cytotoxic stress suggests that this is not the case. However, it is difficult to effectively test this hypothesis without a crystal structure of HSF1 outside of the previously determined DNA binding domain in complex with DNA (Damberger et al 1994, 1995; Littlefield and Nelson 1999). We favor an alternative explanation, in that the LZ3 domain is important for the recruitment of additional factors required for DNA binding in response to stress.…”

Section: Discussionmentioning

confidence: 99%

Genetic Selection for Constitutively Trimerized Human HSF1 Mutants Identifies a Role for Coiled-Coil Motifs in DNA Binding

Neef

Jaeger

Thiele

2013

G3 Genes|Genomes|Genetics

View full text Add to dashboard Cite

Human heat shock transcription factor 1 (HSF1) promotes the expression of stress-responsive genes and is a critical factor for the cellular protective response to proteotoxic and other stresses. In response to stress, HSF1 undergoes a transition from a repressed cytoplasmic monomer to a homotrimer, accumulates in the nucleus, binds DNA, and activates target gene transcription. Although these steps occur as sequential and highly regulated events, our understanding of the full details of the HSF1 activation pathway remains incomplete. Here we describe a genetic screen in humanized yeast that identifies constitutively trimerized HSF1 mutants. Surprisingly, constitutively trimerized HSF1 mutants do not bind to DNA in vivo in the absence of stress and only become DNA binding competent upon stress exposure, suggesting that an additional level of regulation beyond trimerization and nuclear localization may be required for HSF1 DNA binding. Furthermore, we identified a constitutively trimerized and nuclear-localized HSF1 mutant, HSF1 L189P, located in LZ3 of the HSF1 trimerization domain, which in response to proteotoxic stress is strongly compromised for DNA binding at the Hsp70 and Hsp25 promoters but readily binds to the interleukin-6 promoter, suggesting that HSF1 DNA binding is in part regulated in a locus-dependent manner, perhaps via promoter-specific differences in chromatin architecture. Furthermore, these results implicate the LZ3 region of the HSF1 trimerization domain in a function beyond its canonical role in HSF1 trimerization.

show abstract

Refined Solution Structure and Dynamics of the DNA-binding Domain of the Heat Shock Factor fromKluyveromyces lactis

Cited by 20 publications

References 68 publications

The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity

The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity

Structural Classification of HTH DNA-binding Domains and Protein – DNA Interaction Modes

Genetic Selection for Constitutively Trimerized Human HSF1 Mutants Identifies a Role for Coiled-Coil Motifs in DNA Binding

Contact Info

Product

Resources

About