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* Residue conservation analysis
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PDB id:
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Signaling protein
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Title:
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Crystal structure of smad2 mh2 domain bound to the smad-binding domain of sara
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Structure:
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Mad (mothers against decapentaplegic, drosophila) homolog 2. Chain: a, c. Fragment: smad2 mh2 domain. Engineered: yes. Smad anchor for receptor activation. Chain: b, d. Fragment: sara smad2-binding domain. Engineered: yes
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Source:
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Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562. Other_details: this sequence occurs naturally in humans. Other_details: this sequence occurs naturally in humans
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Biol. unit:
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Dimer (from
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Resolution:
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2.20Å
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R-factor:
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0.218
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R-free:
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0.276
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Authors:
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Y.Shi,G.Wu
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Key ref:
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G.Wu
et al.
(2000).
Structural basis of Smad2 recognition by the Smad anchor for receptor activation.
Science,
287,
92-97.
PubMed id:
DOI:
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Date:
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15-Nov-99
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Release date:
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21-Jan-00
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PROCHECK
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Headers
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References
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Q15796
(SMAD2_HUMAN) -
Mothers against decapentaplegic homolog 2 from Homo sapiens
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Seq:
Struc:
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467 a.a.
194 a.a.
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Enzyme class:
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Chains A, B, C, D:
E.C.?
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DOI no:
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Science
287:92-97
(2000)
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PubMed id:
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Structural basis of Smad2 recognition by the Smad anchor for receptor activation.
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G.Wu,
Y.G.Chen,
B.Ozdamar,
C.A.Gyuricza,
P.A.Chong,
J.L.Wrana,
J.Massagué,
Y.Shi.
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ABSTRACT
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The Smad proteins mediate transforming growth factor-beta (TGFbeta) signaling
from the transmembrane serine-threonine receptor kinases to the nucleus. The
Smad anchor for receptor activation (SARA) recruits Smad2 to the TGFbeta
receptors for phosphorylation. The crystal structure of a Smad2 MH2 domain in
complex with the Smad-binding domain (SBD) of SARA has been determined at 2.2
angstrom resolution. SARA SBD, in an extended conformation comprising a rigid
coil, an alpha helix, and a beta strand, interacts with the beta sheet and the
three-helix bundle of Smad2. Recognition between the SARA rigid coil and the
Smad2 beta sheet is essential for specificity, whereas interactions between the
SARA beta strand and the Smad2 three-helix bundle contribute significantly to
binding affinity. Comparison of the structures between Smad2 and a comediator
Smad suggests a model for how receptor-regulated Smads are recognized by the
type I receptors.
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Selected figure(s)
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Figure 2.
Fig. 2. Overall structure of the Smad2 MH2 domain in complex
with a SARA SBD. (A) The schematic representation on the right
panel is related to the one on the left by a 90° rotation
along the horizontal axis. Smad2 and SARA are shown in green and
pink, respectively. The secondary structural elements in SARA
and some prominent features in Smad2 are labeled and
color-coded. (B) Sequence of the SARA SBD showing its secondary
structural elements. The bar graph below sequence shows the
buried surface area per SARA residue upon complex formation. The
residues that are targeted by inactivating mutations are
highlighted in red (Fig. 1C). (C) Superimposition of the
structures of the MH2 domains in Smad2 (green) and Smad4 (red),
shown in stereo view. The disordered loop between helices H3 and
H4 in Smad4 is indicated by a red dotted line. This figure was
prepared with MOLSCRIPT (24).
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Figure 3.
Fig. 3. Schematic representation of the interactions between
Smad2 and SARA. (A) The interactions are predominantly
hydrophobic in nature. The surface of Smad2 MH2 domain is
represented by degrees of hydrophobicity. The C  backbone
of SARA SBD is shown in pink and the buried hydrophobic residues
are highlighted in orange. This figure was prepared with GRASP
(25). (B) A closeup view of the interactions between the rigid
coil of SARA and the strands B8 and B9 of Smad2. Smad2 and SARA
are colored green and pink, respectively. The interacting side
chains are shown in yellow for Smad2 and in purple for SARA. The
O and N atoms are shown as red and blue balls, respectively. The
left panel shows the interface, whereas the right panel shows
the conformation of the rigid coil by itself. Aside from
extensive van der Waals interactions at the interface, there are
a total of five intermolecular H bonds. These include: Ser671 O
 to Tyr366
carbonyl, Pro672 carbonyl to Trp368 N  1, Tyr680
O  to Lys375
amide, Ser682 amide to Asn381 carbonyl, and Ser682 carbonyl to
Asn381 amide. (C) A closeup view of the interactions between the
helix of
SARA and strands B5 and B6 of Smad2. Color coding scheme is
identical to (B). (D) A closeup view of the interactions between
the  strand of
SARA and the three-helix bundle and strand B1' of Smad2.
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The above figures are
reprinted
by permission from the AAAs:
Science
(2000,
287,
92-97)
copyright 2000.
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Figures were
selected
by an automated process.
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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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C.Huang,
R.Du,
P.Zhang,
H.Meng,
H.Jia,
Y.Song,
M.Li,
Y.Zhang,
and
S.Sun
(2011).
Expression, purification, and functional characterization of recombinant PTD-SARA.
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Acta Biochim Biophys Sin (Shanghai),
43,
110-117.
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K.Miyazono,
Y.Kamiya,
and
M.Morikawa
(2010).
Bone morphogenetic protein receptors and signal transduction.
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J Biochem,
147,
35-51.
|
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L.Li,
B.P.Orner,
T.Huang,
A.P.Hinck,
and
L.L.Kiessling
(2010).
Peptide ligands that use a novel binding site to target both TGF-β receptors.
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Mol Biosyst,
6,
2392-2402.
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|
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Y.Watanabe,
S.Itoh,
T.Goto,
E.Ohnishi,
M.Inamitsu,
F.Itoh,
K.Satoh,
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A.Tanaka,
N.Nakano,
A.M.Mommaas,
H.Shibuya,
P.Ten Dijke,
and
M.Kato
(2010).
TMEPAI, a transmembrane TGF-beta-inducible protein, sequesters Smad proteins from active participation in TGF-beta signaling.
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Mol Cell,
37,
123-134.
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C.E.Runyan,
T.Hayashida,
S.Hubchak,
J.F.Curley,
and
H.W.Schnaper
(2009).
Role of SARA (SMAD anchor for receptor activation) in maintenance of epithelial cell phenotype.
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J Biol Chem,
284,
25181-25189.
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|
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C.Wang,
L.Chen,
L.Wang,
and
J.Wu
(2009).
Crystal structure of the MH2 domain of Drosophila Mad.
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Sci China C Life Sci,
52,
539-544.
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PDB code:
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K.Tsuchida,
M.Nakatani,
K.Hitachi,
A.Uezumi,
Y.Sunada,
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and
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Activin signaling as an emerging target for therapeutic interventions.
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Cell Commun Signal,
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M.Fuxreiter,
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A.K.Dunker,
and
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(2009).
Close encounters of the third kind: disordered domains and the interactions of proteins.
|
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Bioessays,
31,
328-335.
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|
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|
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S.Gao,
C.Alarcón,
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S.Rahman,
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P.Tempst,
and
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(2009).
Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta signaling.
|
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Mol Cell,
36,
457-468.
|
|
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|
|
|
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S.I.Kim,
J.H.Kwak,
H.J.Na,
J.K.Kim,
Y.Ding,
and
M.E.Choi
(2009).
Transforming growth factor-beta (TGF-beta1) activates TAK1 via TAB1-mediated autophosphorylation, independent of TGF-beta receptor kinase activity in mesangial cells.
|
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J Biol Chem,
284,
22285-22296.
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R.Hao,
L.Chen,
J.W.Wu,
and
Z.X.Wang
(2008).
Structure of Drosophila Mad MH2 domain.
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Acta Crystallogr Sect F Struct Biol Cryst Commun,
64,
986-990.
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PDB code:
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R.Hariharan,
and
M.R.Pillai
(2008).
Structure-function relationship of inhibitory Smads: Structural flexibility contributes to functional divergence.
|
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Proteins,
71,
1853-1862.
|
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|
|
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S.Ross,
and
C.S.Hill
(2008).
How the Smads regulate transcription.
|
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Int J Biochem Cell Biol,
40,
383-408.
|
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|
|
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|
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T.Mori,
K.Kitano,
S.Terawaki,
R.Maesaki,
Y.Fukami,
and
T.Hakoshima
(2008).
Structural basis for CD44 recognition by ERM proteins.
|
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J Biol Chem,
283,
29602-29612.
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PDB code:
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F.Dai,
C.Chang,
X.Lin,
P.Dai,
L.Mei,
and
X.H.Feng
(2007).
Erbin inhibits transforming growth factor beta signaling through a novel Smad-interacting domain.
|
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Mol Cell Biol,
27,
6183-6194.
|
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|
|
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M.E.Hahn,
J.P.Pellois,
M.Vila-Perelló,
and
T.W.Muir
(2007).
Tunable photoactivation of a post-translationally modified signaling protein and its unmodified counterpart in live cells.
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Chembiochem,
8,
2100-2105.
|
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P.T.Loverde,
A.Osman,
and
A.Hinck
(2007).
Schistosoma mansoni: TGF-beta signaling pathways.
|
| |
Exp Parasitol,
117,
304-317.
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M.Kowanetz,
H.Sugino,
Y.Yoneda,
C.H.Heldin,
and
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(2006).
The mechanism of nuclear export of Smad3 involves exportin 4 and Ran.
|
| |
Mol Cell Biol,
26,
1318-1332.
|
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|
|
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|
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B.M.Zhao,
and
F.M.Hoffmann
(2006).
Inhibition of transforming growth factor-beta1-induced signaling and epithelial-to-mesenchymal transition by the Smad-binding peptide aptamer Trx-SARA.
|
| |
Mol Biol Cell,
17,
3819-3831.
|
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H.Remaut,
and
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Protein-protein interaction through beta-strand addition.
|
| |
Trends Biochem Sci,
31,
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W.Pirovano,
K.A.Feenstra,
and
J.Heringa
(2006).
Sequence comparison by sequence harmony identifies subtype-specific functional sites.
|
| |
Nucleic Acids Res,
34,
6540-6548.
|
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B.Y.Qin,
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H.Srinath,
S.S.Lam,
J.J.Correia,
R.Derynck,
and
K.Lin
(2005).
Crystal structure of IRF-3 in complex with CBP.
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| |
Structure,
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and
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| |
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B.Zhao,
and
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(2005).
Selective inhibition of TGF-beta responsive genes by Smad-interacting peptide aptamers from FoxH1, Lef1 and CBP.
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| |
Oncogene,
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3864-3874.
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and
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S.Hashimoto,
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and
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Systematic identification of human melanoma antigens using serial analysis of gene expression (SAGE).
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J Immunother (1997),
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C.Le Roy,
and
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(2004).
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Nature,
431,
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L.Xu,
and
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(2004).
Nucleocytoplasmic shuttling of signal transducers.
|
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Nat Rev Mol Cell Biol,
5,
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|
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|
|
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M.Kondo,
H.Suzuki,
K.Takehara,
K.Miyazono,
and
M.Kato
(2004).
Transforming growth factor-beta signaling is differentially inhibited by Smad2D450E and Smad3D407E.
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| |
Cancer Sci,
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M.de Caestecker
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C.S.Page,
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and
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Recognition of phosphorylated-Smad2-containing complexes by a novel Smad interaction motif.
|
| |
Mol Cell Biol,
24,
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|
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R.S.Peterson,
R.A.Andhare,
K.T.Rousche,
W.Knudson,
W.Wang,
J.B.Grossfield,
R.O.Thomas,
R.E.Hollingsworth,
and
C.B.Knudson
(2004).
CD44 modulates Smad1 activation in the BMP-7 signaling pathway.
|
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J Cell Biol,
166,
1081-1091.
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B.Y.Qin,
C.Liu,
S.S.Lam,
H.Srinath,
R.Delston,
J.J.Correia,
R.Derynck,
and
K.Lin
(2003).
Crystal structure of IRF-3 reveals mechanism of autoinhibition and virus-induced phosphoactivation.
|
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Nat Struct Biol,
10,
913-921.
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PDB code:
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T.Reguly,
and
J.L.Wrana
(2003).
In or out? The dynamics of Smad nucleocytoplasmic shuttling.
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Trends Cell Biol,
13,
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Y.Shi,
and
J.Massagué
(2003).
Mechanisms of TGF-beta signaling from cell membrane to the nucleus.
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| |
Cell,
113,
685-700.
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A.Mehra,
and
J.L.Wrana
(2002).
TGF-beta and the Smad signal transduction pathway.
|
| |
Biochem Cell Biol,
80,
605-622.
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B.Y.Qin,
S.S.Lam,
J.J.Correia,
and
K.Lin
(2002).
Smad3 allostery links TGF-beta receptor kinase activation to transcriptional control.
|
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Genes Dev,
16,
1950-1963.
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PDB codes:
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D.U.Kloos,
C.Choi,
and
E.Wingender
(2002).
The TGF-beta--Smad network: introducing bioinformatic tools.
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S.Itoh,
and
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(2002).
The FYVE domain in Smad anchor for receptor activation (SARA) is sufficient for localization of SARA in early endosomes and regulates TGF-beta/Smad signalling.
|
| |
Genes Cells,
7,
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J.L.Wrana
(2002).
Phosphoserine-dependent regulation of protein-protein interactions in the Smad pathway.
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| |
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S.Cöl,
and
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Smad2 nucleocytoplasmic shuttling by nucleoporins CAN/Nup214 and Nup153 feeds TGFbeta signaling complexes in the cytoplasm and nucleus.
|
| |
Mol Cell,
10,
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|
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A.Hatzubai,
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and
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F.Itoh,
and
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| |
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|
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P.A.Bates,
and
C.S.Hill
(2002).
Different Smad2 partners bind a common hydrophobic pocket in Smad2 via a defined proline-rich motif.
|
| |
EMBO J,
21,
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| |
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R.G.Phelps,
H.Spiera,
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(2002).
Halofuginone, an inhibitor of type-I collagen synthesis and skin sclerosis, blocks transforming-growth-factor-beta-mediated Smad3 activation in fibroblasts.
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| |
J Invest Dermatol,
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M.P.de Caestecker,
J.J.Correia,
and
K.Lin
(2001).
Structural basis of Smad1 activation by receptor kinase phosphorylation.
|
| |
Mol Cell,
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1303-1312.
|
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PDB code:
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C.Prunier,
N.Ferrand,
B.Frottier,
M.Pessah,
and
A.Atfi
(2001).
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| |
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J.Rich,
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and
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(2001).
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|
| |
Microsc Res Tech,
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|
J.W.Wu,
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J.Seoane,
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T.W.Muir,
R.Fairman,
J.Massagué,
and
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(2001).
Crystal structure of a phosphorylated Smad2. Recognition of phosphoserine by the MH2 domain and insights on Smad function in TGF-beta signaling.
|
| |
Mol Cell,
8,
1277-1289.
|
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PDB code:
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J.Yue,
and
K.M.Mulder
(2001).
Transforming growth factor-beta signal transduction in epithelial cells.
|
| |
Pharmacol Ther,
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|
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|
L.Attisano,
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(2001).
The Smads.
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Genome Biol,
2,
REVIEWS3010.
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M.Furuhashi,
K.Yagi,
H.Yamamoto,
Y.Furukawa,
S.Shimada,
Y.Nakamura,
A.Kikuchi,
K.Miyazono,
and
M.Kato
(2001).
Axin facilitates Smad3 activation in the transforming growth factor beta signaling pathway.
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Mol Cell Biol,
21,
5132-5141.
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M.Huse,
T.W.Muir,
L.Xu,
Y.G.Chen,
J.Kuriyan,
and
J.Massagué
(2001).
The TGF beta receptor activation process: an inhibitor- to substrate-binding switch.
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Mol Cell,
8,
671-682.
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PDB code:
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Y.Shi
(2001).
Structural insights on Smad function in TGFbeta signaling.
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Bioessays,
23,
223-232.
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D.Durocher,
I.A.Taylor,
D.Sarbassova,
L.F.Haire,
S.L.Westcott,
S.P.Jackson,
S.J.Smerdon,
and
M.B.Yaffe
(2000).
The molecular basis of FHA domain:phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms.
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| |
Mol Cell,
6,
1169-1182.
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PDB code:
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J.L.Wrana
(2000).
Regulation of Smad activity.
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Cell,
100,
189-192.
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J.L.Wrana,
and
L.Attisano
(2000).
The Smad pathway.
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Cytokine Growth Factor Rev,
11,
5.
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J.Massagué,
and
D.Wotton
(2000).
Transcriptional control by the TGF-beta/Smad signaling system.
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EMBO J,
19,
1745-1754.
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L.Attisano,
and
J.L.Wrana
(2000).
Smads as transcriptional co-modulators.
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| |
Curr Opin Cell Biol,
12,
235-243.
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S.A.Pangas,
and
T.K.Woodruff
(2000).
Activin signal transduction pathways.
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| |
Trends Endocrinol Metab,
11,
309-314.
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S.Itoh,
F.Itoh,
M.J.Goumans,
and
P.Ten Dijke
(2000).
Signaling of transforming growth factor-beta family members through Smad proteins.
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Eur J Biochem,
267,
6954-6967.
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The most recent references are shown first.
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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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