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PDBsum entry 2bf0
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Transcription
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PDB id
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2bf0
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References listed in PDB file
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Key reference
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Title
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Key features of the interaction between pcf11 cid and RNA polymerase ii ctd.
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Authors
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C.G.Noble,
D.Hollingworth,
S.R.Martin,
V.Ennis-Adeniran,
S.J.Smerdon,
G.Kelly,
I.A.Taylor,
A.Ramos.
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Ref.
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Nat Struct Mol Biol, 2005,
12,
144-151.
[DOI no: ]
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PubMed id
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Abstract
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The C-terminal domain (CTD) of the large subunit of RNA polymerase II is a
platform for mRNA processing factors and links gene transcription to mRNA
capping, splicing and polyadenylation. Pcf11, an essential component of the mRNA
cleavage factor IA, contains a CTD-interaction domain that binds in a
phospho-dependent manner to the heptad repeats within the RNA polymerase II CTD.
We show here that the phosphorylated CTD exists as a dynamic disordered ensemble
in solution and, by induced fit, it assumes a structured conformation when bound
to Pcf11. In addition, we detected cis-trans populations for the CTD prolines,
and found that only the all-trans form is selected for binding. These data
suggest that the recognition of the CTD is regulated by independent
site-specific modifications (phosphorylation and proline cis-trans
isomerization) and, probably, by the local concentration of suitable binding
sites.
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Figure 1.
Figure 1. Peptides used in this study. In the table (bottom)
the first amino acid of each peptide has been aligned to the
corresponding position within the first repeat (repeat a). The
nomenclature used in the manuscript identifies (i) the amino
acid type (ii) its number within the repeat, and (iii) the
repeat it belongs to. Phosphorylated residues are indicated in
red. A graphical representation of one heptad repeat is shown.
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Figure 6.
Figure 6. Binding of Pcf11 CID to CTD-derived peptides. (a)
1H-decoupled 1D 31P NMR spectra of YSpPTSPS (top); (YSpPTSPS)[2]
(middle); and (YSpPTSPS)[3] (bottom). The downfield peak in the
spectrum can be assigned to the 31P resonance of Ser[2a] (S[2a])
as this peak does not change substantially in the three spectra.
(b) 1H-decoupled 1D 31P NMR spectra recorded on (YSpPTSPS)[3],
alone and with increasing concentration of Pcf11(1 -142) (ratios
of 1:0, 1:1 and 1:3, top to bottom). The 31P resonances of
Ser[2b] and Ser[2c] (S[2b] and S[2c]) broaden and shift upon
protein binding, whereas Ser[2a] (S[2a]) is essentially
identical in the free and bound form. (c) Chemical shift
perturbation of Pcf11 CID upon binding to (YSpPTSPS)[2] (top)
and (YSpPTSPS)[3] (bottom). The weighted chemical shift changes
are plotted against the CID sequence. Well-dispersed resonances
in the free protein spectrum that could not be assigned in the
bound protein spectrum were given a weighted chemical shift
change value of 0.5 as default.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nat Struct Mol Biol
(2005,
12,
144-151)
copyright 2005.
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