Figure 6.
Figure 6 Effect of CTD conformation on functional activity of
Gfh1. (A) Model structures of mutant factors with conformations
fixed via S–S bridges, Gfh1-CC12 and Gfh1-CC13 are shown as
ribbons. Models were generated by Swiss-Model (Schwede et al,
2003) using the structures of Tth Gfh1 and E. coli GreA as
templates, respectively. (B) Summary of the inhibitory
activities of wt and mutant Gfh1-CC factors. The IC[50] values
were obtained from abortive initiation assay as in Figure
4A–C, conducted under indicated conditions. (C)
[^32P]Gfh1–RNAP competition-binding assay. [^32P]Gfh1–RNAP
core complex was chromatographed with or without 20 M
competitor proteins, Gfh1-CC12 or Gfh1-CC13, at pH 6.4 under
nonreducing conditions (see Figure 4D). Free oxidized forms of
Gfh1-CC12 and Gfh1-CC13 all elute irrespective of pH with almost
identical retention times of 24.5–24.7 min (the same as that
of the wt Gfh1) corresponding to an apparent molecular weight of
26
kDa (data not shown), which, according to a light-scattering
analysis, represents a monomer (see Supplementary data).
The above figure is reprinted
by permission from Macmillan Publishers Ltd:
EMBO J
(2006,
25,
2131-2141)
copyright 2006.
M
competitor proteins, Gfh1-CC12 or Gfh1-CC13, at pH 6.4 under
nonreducing conditions (see Figure 4D). Free oxidized forms of
Gfh1-CC12 and Gfh1-CC13 all elute irrespective of pH with almost
identical retention times of 24.5–24.7 min (the same as that
of the wt Gfh1) corresponding to an apparent molecular weight of
26
kDa (data not shown), which, according to a light-scattering
analysis, represents a monomer (see Supplementary data).