PDBsum entry 2j6y

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protein Protein-protein interface(s) links
Hydrolase PDB id
Protein chains
85 a.a. *
Waters ×348
* Residue conservation analysis
PDB id:
Name: Hydrolase
Title: Structural and functional characterisation of partner switching regulating the environmental stress response in bacillus subtilis
Structure: Phosphoserine phosphatase rsbu. Chain: a, b, c, d, e. Fragment: rsbt binding domain, residues 1-111. Synonym: sigma factor sigb regulation protein rsbu, n-rsbu. Engineered: yes. Mutation: yes
Source: Bacillus subtilis. Organism_taxid: 1423. Expressed in: escherichia coli. Expression_system_taxid: 511693. Expression_system_variant: de3.
1.85Å     R-factor:   0.186     R-free:   0.253
Authors: S.W.Hardwick,J.Pane-Farre,O.Delumeau,J.Marles-Wright, J.W.Murray,M.Hecker,R.J.Lewis
Key ref:
S.W.Hardwick et al. (2007). Structural and functional characterization of partner switching regulating the environmental stress response in Bacillus subtilis. J Biol Chem, 282, 11562-11572. PubMed id: 17303566 DOI: 10.1074/jbc.M609733200
05-Oct-06     Release date:   13-Feb-07    
Go to PROCHECK summary

Protein chains
Pfam   ArchSchema ?
P40399  (RSBU_BACSU) -  Phosphoserine phosphatase RsbU
335 a.a.
85 a.a.*
Key:    PfamA domain  Secondary structure  CATH domain
* PDB and UniProt seqs differ at 1 residue position (black cross)

 Enzyme reactions 
   Enzyme class: E.C.  - Phosphoserine phosphatase.
[IntEnz]   [ExPASy]   [KEGG]   [BRENDA]
      Reaction: O-phospho-L(or D)-serine + H2O = L(or D)-serine + phosphate
O-phospho-L(or D)-serine
+ H(2)O
= L(or D)-serine
+ phosphate
Molecule diagrams generated from .mol files obtained from the KEGG ftp site
 Gene Ontology (GO) functional annotation 
  GO annot!
  Biological process     protein phosphorylation   1 term 


    Key reference    
DOI no: 10.1074/jbc.M609733200 J Biol Chem 282:11562-11572 (2007)
PubMed id: 17303566  
Structural and functional characterization of partner switching regulating the environmental stress response in Bacillus subtilis.
S.W.Hardwick, J.Pané-Farré, O.Delumeau, J.Marles-Wright, J.W.Murray, M.Hecker, R.J.Lewis.
The general stress response of Bacillus subtilis and close relatives provides the cell with protection from a variety of stresses. The upstream component of the environmental stress signal transduction cascade is activated by the RsbT kinase that switches binding partners from a 25 S macromolecular complex, the stressosome, to the RsbU phosphatase. Once the RsbU phosphatase is activated by interacting with RsbT, the alternative sigma factor, sigmaB, directs transcription of the general stress regulon. Previously, we demonstrated that the N-terminal domain of RsbU mediates the binding of RsbT. We now describe residues in N-RsbU that are crucial to this interaction by experimentation both in vitro and in vivo. Furthermore, crystal structures of the N-RsbU mutants provide a molecular explanation for the loss of interaction. Finally, we also characterize mutants in RsbT that affect binding to both RsbU and a simplified, binary model of the stressosome and thus identify overlapping binding surfaces on the RsbT "switch."
  Selected figure(s)  
Figure 3.
FIGURE 3. Crystallographic analysis of N-RsbU mutants. a, molecular surface of the N-RsbU dimer is colored mostly blue. Surfaces colored red correspond to residues targeted in this study as potential RsbT-interacting partners and are also conserved across RsbT-encoding bacterial species; residues colored green are conserved irrespective of the presence of rsbT in the genome, and the orange line delineates the surface region of those amino acids that this study has revealed to mediate RsbT binding in RsbU. The yellow surface represents Ser^61, away from the proposed RsbT-binding surface. Primes are used to indicate that the residue in question comes from the 2nd protomer in the N-RsbU dimer. b-d, OMIT electron density maps (each contoured at a level of 1 ) associated with the structures of the N-RsbU mutants E24K (b), Y28I (c), and I78K (d). The final models are shown as solid sticks with blue carbon atoms and with the wild type structure at the mutated position with yellow carbon atoms. For E24K, four of the five independent copies of N-RsbU in the asymmetric unit have very similar side chain conformations at position 24. In all instances, the rotamer of the introduced side chain is one of the two most frequently observed conformations in the COOT rotamer library (38). Ribbon schematics of N-RsbU (e) and N-RsbR (f) colored blue and green, respectively; highlighted in red are the positions of mutations in both proteins that disrupt the binding of RsbT, indicating that a common surface is used.
Figure 5.
FIGURE 5. Schematic of the environmental stress response. Pre-stress, the RsbT kinase is sequestered by the stressosome, represented here as a round disk with peripheral turrets. On the imposition of stress, the kinase activity of RsbT results in the phosphorylation of the stressosome, and RsbT is released to interact with RsbU and stimulate its phosphatase function toward RsbV-P. Dephosphorylated RsbV attacks the RsbW- ^B complex and liberates ^B to bind to the crab claw-shaped core RNA polymerase (cRNAP) to form the holoenzyme (hRNAP) that then transcribes the ^B regulon. To reset the switch, the RsbX phosphatase must dephosphorylate the phosphorylated stressosome, which can then re-capture RsbT, down-regulating RsbU phosphatase activity. The kinase function of RsbW then predominates, and RsbV is maintained in a phosphorylated form, and ^B is retained by RsbW. In the scheme, enzymatic activities are italicized, and partner switching modules linked by dashed arrows.
  The above figures are reprinted by permission from the ASBMB: J Biol Chem (2007, 282, 11562-11572) copyright 2007.  
  Figures were selected by the author.  

Literature references that cite this PDB file's key reference

  PubMed id Reference
21220116 J.King-Scott, P.V.Konarev, S.Panjikar, R.Jordanova, D.I.Svergun, and P.A.Tucker (2011).
Structural characterization of the multidomain regulatory protein Rv1364c from Mycobacterium tuberculosis.
  Structure, 19, 56-69.  
21264304 N.Ondrusch, and J.Kreft (2011).
Blue and red light modulates SigB-dependent gene transcription, swimming motility and invasiveness in Listeria monocytogenes.
  PLoS One, 6, e16151.  
20019076 A.Reeves, L.Martinez, and W.Haldenwang (2010).
Expression of, and in vivo stressosome formation by, single members of the RsbR protein family in Bacillus subtilis.
  Microbiology, 156, 990-998.  
20233929 Q.C.Truong-Bolduc, and D.C.Hooper (2010).
Phosphorylation of MgrA and its effect on expression of the NorA and NorB efflux pumps of Staphylococcus aureus.
  J Bacteriol, 192, 2525-2534.  
19638175 J.Klebensberger, A.Birkenmaier, R.Geffers, S.Kjelleberg, and B.Philipp (2009).
SiaA and SiaD are essential for inducing autoaggregation as a specific response to detergent stress in Pseudomonas aeruginosa.
  Environ Microbiol, 11, 3073-3086.  
19201800 J.Pané-Farré, B.Jonas, S.W.Hardwick, K.Gronau, R.J.Lewis, M.Hecker, and S.Engelmann (2009).
Role of RsbU in controlling SigB activity in Staphylococcus aureus following alkaline stress.
  J Bacteriol, 191, 2561-2573.  
  19704888 J.Marles-Wright, and R.J.Lewis (2008).
The Bacillus subtilis stressosome: A signal integration and transduction hub.
  Commun Integr Biol, 1, 182-184.  
18832644 J.Marles-Wright, T.Grant, O.Delumeau, G.van Duinen, S.J.Firbank, P.J.Lewis, J.W.Murray, J.A.Newman, M.B.Quin, P.R.Race, A.Rohou, W.Tichelaar, M.van Heel, and R.J.Lewis (2008).
Molecular architecture of the "stressosome," a signal integration and transduction hub.
  Science, 322, 92-96.
PDB code: 2vy9
19003856 M.Hecker, H.Antelmann, K.Büttner, and J.Bernhardt (2008).
Gel-based proteomics of Gram-positive bacteria: a powerful tool to address physiological questions.
  Proteomics, 8, 4958-4975.  
18805983 Q.C.Truong-Bolduc, Y.Ding, and D.C.Hooper (2008).
Posttranslational modification influences the effects of MgrA on norA expression in Staphylococcus aureus.
  J Bacteriol, 190, 7375-7381.  
17920859 J.Marles-Wright, and R.J.Lewis (2007).
Stress responses of bacteria.
  Curr Opin Struct Biol, 17, 755-760.  
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. Where a reference describes a PDB structure, the PDB code is shown on the right.