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PDBsum entry 4qht
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Transcription
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PDB id
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4qht
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PDB id:
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| Name: |
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Transcription
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Title:
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Crystal structure of aaa+/ sigma 54 activator domain of the flagellar regulatory protein flrc from vibrio cholerae in atp analog bound state
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Structure:
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Flagellar regulatory protein c. Chain: a, b, c, d, e, f, g. Fragment: unp residues 132-381. Engineered: yes
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Source:
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Vibrio cholerae. Organism_taxid: 345073. Strain: o395. Gene: flrc. Expressed in: escherichia coli. Expression_system_taxid: 562.
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Resolution:
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2.56Å
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R-factor:
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0.186
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R-free:
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0.247
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Authors:
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S.Dey,M.Biswas,U.Sen,J.Dasgupta
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Key ref:
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S.Dey
et al.
(2015).
Unique ATPase site architecture triggers cis-mediated synchronized ATP binding in heptameric AAA+-ATPase domain of flagellar regulatory protein FlrC.
J Biol Chem,
290,
8734-8747.
PubMed id:
DOI:
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Date:
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29-May-14
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Release date:
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16-Jul-14
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PROCHECK
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Headers
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References
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A0A0H3AHP1
(A0A0H3AHP1_VIBC3) -
Flagellar regulatory protein C from Vibrio cholerae serotype O1 (strain ATCC 39541 / Classical Ogawa 395 / O395)
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Seq:
Struc:
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479 a.a.
246 a.a.*
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Key: |
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PfamA domain |
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Secondary structure |
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CATH domain |
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*
PDB and UniProt seqs differ
at 2 residue positions (black
crosses)
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DOI no:
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J Biol Chem
290:8734-8747
(2015)
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PubMed id:
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Unique ATPase site architecture triggers cis-mediated synchronized ATP binding in heptameric AAA+-ATPase domain of flagellar regulatory protein FlrC.
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S.Dey,
M.Biswas,
U.Sen,
J.Dasgupta.
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ABSTRACT
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Bacterial enhancer-binding proteins (bEBPs) oligomerize through AAA(+) domains
and use ATP hydrolysis-driven energy to isomerize the RNA polymerase-σ(54)
complex during transcriptional initiation. Here, we describe the first structure
of the central AAA(+) domain of the flagellar regulatory protein FlrC (FlrC(C)),
a bEBP that controls flagellar synthesis in Vibrio cholerae. Our results showed
that FlrC(C) forms heptamer both in nucleotide (Nt)-free and -bound states
without ATP-dependent subunit remodeling. Unlike the bEBPs such as NtrC1 or
PspF, a novel cis-mediated "all or none" ATP binding occurs in the
heptameric FlrC(C), because constriction at the ATPase site, caused by loop L3
and helix α7, restricts the proximity of the trans-protomer required for Nt
binding. A unique "closed to open" movement of Walker A, assisted by
trans-acting "Glu switch" Glu-286, facilitates ATP binding and
hydrolysis. Fluorescence quenching and ATPase assays on FlrC(C) and mutants
revealed that although Arg-349 of sensor II, positioned by trans-acting Glu-286
and Tyr-290, acts as a key residue to bind and hydrolyze ATP, Arg-319 of α7
anchors ribose and controls the rate of ATP hydrolysis by retarding the
expulsion of ADP. Heptameric state of FlrC(C) is restored in solution even with
the transition state mimicking ADP·AlF3. Structural results and pulldown assays
indicated that L3 renders an in-built geometry to L1 and L2 causing
σ(54)-FlrC(C) interaction independent of Nt binding. Collectively, our results
underscore a novel mechanism of ATP binding and σ(54) interaction that strives
to understand the transcriptional mechanism of the bEBPs, which probably
interact directly with the RNA polymerase-σ(54) complex without DNA looping.
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