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PDBsum entry 2eeu
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Biochemistry
46:13297-13309
(2007)
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PubMed id:
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Mutational analysis of the purine riboswitch aptamer domain.
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S.D.Gilbert,
C.E.Love,
A.L.Edwards,
R.T.Batey.
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ABSTRACT
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The purine riboswitch is one of a number of mRNA elements commonly found in the
5'-untranslated region capable of controlling expression in a cis-fashion via
its ability to directly bind small-molecule metabolites. Extensive biochemical
and structural analysis of the nucleobase-binding domain of the riboswitch,
referred to as the aptamer domain, has revealed that the mRNA recognizes its
cognate ligand using an intricately folded three-way junction motif that
completely encapsulates the ligand. High-affinity binding of the purine
nucleobase is facilitated by a distal loop-loop interaction that is conserved
between both the adenine and guanine riboswitches. To understand the
contribution of conserved nucleotides in both the three-way junction and the
loop-loop interaction of this RNA, we performed a detailed mutagenic survey of
these elements in the context of an adenine-responsive variant of the xpt-pbuX
guanine riboswitch from Bacillus subtilis. The varying ability of these mutants
to bind ligand as measured by isothermal titration calorimetry uncovered the
conserved nucleotides whose identity is required for purine binding.
Crystallographic analysis of the bound form of five mutants and chemical probing
of their free state demonstrate that the identity of several universally
conserved nucleotides is not essential for formation of the RNA-ligand complex
but rather for maintaining a binding-competent form of the free RNA. These data
show that conservation patterns in riboswitches arise from a combination of
formation of the ligand-bound complex, promoting an open form of the free RNA,
and participating in the secondary structural switch with the expression
platform.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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B.Heppell,
S.Blouin,
A.M.Dussault,
J.Mulhbacher,
E.Ennifar,
J.C.Penedo,
and
D.A.Lafontaine
(2011).
Molecular insights into the ligand-controlled organization of the SAM-I riboswitch.
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Nat Chem Biol,
7,
384-392.
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J.Grojean,
and
B.Downes
(2010).
Riboswitches as hormone receptors: hypothetical cytokinin-binding riboswitches in Arabidopsis thaliana.
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Biol Direct,
5,
60.
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K.S.Keating,
and
A.M.Pyle
(2010).
Semiautomated model building for RNA crystallography using a directed rotameric approach.
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Proc Natl Acad Sci U S A,
107,
8177-8182.
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M.D.Brenner,
M.S.Scanlan,
M.K.Nahas,
T.Ha,
and
S.K.Silverman
(2010).
Multivector fluorescence analysis of the xpt guanine riboswitch aptamer domain and the conformational role of guanine.
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Biochemistry,
49,
1596-1605.
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N.J.Baird,
and
A.R.Ferré-D'Amaré
(2010).
Idiosyncratically tuned switching behavior of riboswitch aptamer domains revealed by comparative small-angle X-ray scattering analysis.
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RNA,
16,
598-609.
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V.Delfosse,
P.Bouchard,
E.Bonneau,
P.Dagenais,
J.F.Lemay,
D.A.Lafontaine,
and
P.Legault
(2010).
Riboswitch structure: an internal residue mimicking the purine ligand.
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Nucleic Acids Res,
38,
2057-2068.
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PDB code:
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A.D.Garst,
and
R.T.Batey
(2009).
A switch in time: detailing the life of a riboswitch.
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Biochim Biophys Acta,
1789,
584-591.
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A.L.Edwards,
and
R.T.Batey
(2009).
A structural basis for the recognition of 2'-deoxyguanosine by the purine riboswitch.
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J Mol Biol,
385,
938-948.
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PDB code:
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A.Serganov
(2009).
The long and the short of riboswitches.
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Curr Opin Struct Biol,
19,
251-259.
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M.Sharma,
G.Bulusu,
and
A.Mitra
(2009).
MD simulations of ligand-bound and ligand-free aptamer: molecular level insights into the binding and switching mechanism of the add A-riboswitch.
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RNA,
15,
1673-1692.
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M.de la Peña,
D.Dufour,
and
J.Gallego
(2009).
Three-way RNA junctions with remote tertiary contacts: a recurrent and highly versatile fold.
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RNA,
15,
1949-1964.
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O.Prychyna,
M.S.Dahabieh,
J.Chao,
and
M.A.O'Neill
(2009).
Sequence-dependent folding and unfolding of ligand-bound purine riboswitches.
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Biopolymers,
91,
953-965.
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P.Sharma,
S.Sharma,
M.Chawla,
and
A.Mitra
(2009).
Modeling the noncovalent interactions at the metabolite binding site in purine riboswitches.
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J Mol Model,
15,
633-649.
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S.D.Gilbert,
F.E.Reyes,
A.L.Edwards,
and
R.T.Batey
(2009).
Adaptive ligand binding by the purine riboswitch in the recognition of guanine and adenine analogs.
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Structure,
17,
857-868.
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PDB codes:
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T.H.Chang,
H.D.Huang,
L.C.Wu,
C.T.Yeh,
B.J.Liu,
and
J.T.Horng
(2009).
Computational identification of riboswitches based on RNA conserved functional sequences and conformations.
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RNA,
15,
1426-1430.
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C.D.Stoddard,
S.D.Gilbert,
and
R.T.Batey
(2008).
Ligand-dependent folding of the three-way junction in the purine riboswitch.
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RNA,
14,
675-684.
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T.H.Chang,
J.T.Horng,
and
H.D.Huang
(2008).
RNALogo: a new approach to display structural RNA alignment.
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Nucleic Acids Res,
36,
W91-W96.
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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.
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