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PDBsum entry 1fbu

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protein Protein-protein interface(s) links
Transcription PDB id
1fbu
Jmol
Contents
Protein chains
89 a.a. *
Waters ×129
* Residue conservation analysis
PDB id:
1fbu
Name: Transcription
Title: Heat shock transcription factor DNA binding domain
Structure: Heat shock factor protein. Chain: a, b. Fragment: hsf DNA binding domain. Engineered: yes
Source: Kluyveromyces lactis. Organism_taxid: 28985. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
Biol. unit: Not given
Resolution:
2.00Å     R-factor:   0.226     R-free:   0.267
Authors: J.A.Hardy,H.C.M.Nelson
Key ref: J.A.Hardy and H.C.Nelson (2000). Proline in alpha-helical kink is required for folding kinetics but not for kinked structure, function, or stability of heat shock transcription factor. Protein Sci, 9, 2128-2141. PubMed id: 11305238 DOI: 10.1110/ps.9.11.2128
Date:
16-Jul-00     Release date:   10-Jan-01    
PROCHECK
Go to PROCHECK summary
 Headers
 References

Protein chains
Pfam   ArchSchema ?
P22121  (HSF_KLULA) -  Heat shock factor protein
Seq:
Struc:
 
Seq:
Struc:
677 a.a.
89 a.a.*
Key:    PfamA domain  PfamB domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 2 residue positions (black crosses)

 Gene Ontology (GO) functional annotation 
  GO annot!
  Cellular component     nucleus   1 term 
  Biological process     regulation of transcription, DNA-dependent   1 term 
  Biochemical function     sequence-specific DNA binding transcription factor activity     2 terms  

 

 
DOI no: 10.1110/ps.9.11.2128 Protein Sci 9:2128-2141 (2000)
PubMed id: 11305238  
 
 
Proline in alpha-helical kink is required for folding kinetics but not for kinked structure, function, or stability of heat shock transcription factor.
J.A.Hardy, H.C.Nelson.
 
  ABSTRACT  
 
The DNA-binding domain of the yeast heat shock transcription factor (HSF) contains a strictly conserved proline that is at the center of a kink. To define the role of this conserved proline-centered kink, we replaced the proline with a number of other residues. These substitutions did not diminish the ability of the full-length protein to support growth of yeast or to activate transcription, suggesting that the proline at the center of the kink is not conserved for function. The stability of the isolated mutant DNA-binding domains was unaltered from the wild-type, so the proline is not conserved to maintain the stability of the protein. The crystal structures of two of the mutant DNA-binding domains revealed that the helices in the mutant proteins were still kinked after substitution of the proline, suggesting that the proline does not cause the alpha-helical kink. So why are prolines conserved in this and the majority of other kinked alpha-helices if not for structure, function, or stability? The mutant DNA-binding domains are less soluble than wild-type when overexpressed. In addition, the folding kinetics, as measured by stopped-flow fluorescence, is faster for the mutant proteins. These two results support the premise that the presence of the proline is critical for the folding pathway of HSF's DNA-binding domain. The finding may also be more general and explain why kinked helices maintain their prolines.
 

Literature references that cite this PDB file's key reference

  PubMed id Reference
20095047 C.Andrésen, S.Jalal, D.Aili, Y.Wang, S.Islam, A.Jarl, B.Liedberg, B.Wretlind, L.G.Mårtensson, and M.Sunnerhagen (2010).
Critical biophysical properties in the Pseudomonas aeruginosa efflux gene regulator MexR are targeted by mutations conferring multidrug resistance.
  Protein Sci, 19, 680-692.  
16428449 J.Anckar, V.Hietakangas, K.Denessiouk, D.J.Thiele, M.S.Johnson, and L.Sistonen (2006).
Inhibition of DNA binding by differential sumoylation of heat shock factors.
  Mol Cell Biol, 26, 955-964.  
16936821 S.R.Uffenbeck, and J.E.Krebs (2006).
The role of chromatin structure in regulating stress-induced transcription in Saccharomyces cerevisiae.
  Biochem Cell Biol, 84, 477-489.  
16980499 T.R.Raghunand, and W.R.Bishai (2006).
Mapping essential domains of Mycobacterium smegmatis WhmD: insights into WhiB structure and function.
  J Bacteriol, 188, 6966-6976.  
15313242 A.Senes, D.E.Engel, and W.F.DeGrado (2004).
Folding of helical membrane proteins: the role of polar, GxxxG-like and proline motifs.
  Curr Opin Struct Biol, 14, 465-479.  
14997553 Z.Kovári, and M.Vas (2004).
Protein conformer selection by sequence-dependent packing contacts in crystals of 3-phosphoglycerate kinase.
  Proteins, 55, 198-209.  
11406580 M.M.Krem, and E.Di Cera (2001).
Molecular markers of serine protease evolution.
  EMBO J, 20, 3036-3045.  
11292844 M.P.Cicero, S.T.Hubl, C.J.Harrison, O.Littlefield, J.A.Hardy, and H.C.Nelson (2001).
The wing in yeast heat shock transcription factor (HSF) DNA-binding domain is required for full activity.
  Nucleic Acids Res, 29, 1715-1723.  
The most recent references are shown first. Citation data come partly from CiteXplore and partly from an automated harvesting procedure. Note that this is likely to be only a partial list as not all journals are covered by either method. However, we are continually building up the citation data so more and more references will be included with time.