1. Molecular Structure of Human TFIIH
Patrick Schultz, Sébastien Fribourg, Arnaud Poterszman, Véronique Mallouh, Dino Moras, Jean Marc Egly 
Cell 
Volume 102, Issue 5, Pages (September 2000)
DOI: /S (00)

2. Figure 1
Subunit Composition and Transcriptional Activity of TFIIH and rIIH5
(A) The purity and subunit composition of TFIIH and rIIH5 were analyzed by SDS-PAGE and Coomassie blue staining. (*) stands for a contaminant previously identified as eIF2 (Marinoni et al. 1997). (B) In vitro transcription assay was performed with TFIIH and rIIH5 as described (Gerard et al. 1991). Equal amounts of purified CAK/XPD and rIIH5 were mixed for the assay.
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3. Figure 2
Electron Microscopy Observation and Image Analysis of the Human TFIIH Complex
(A) Electron micrograph of TFIIH molecules adsorbed on a carbon film and negatively stained with uranyl acetate showing the homogeneity in size and the dispersion of the complexes. (B) Gallery of the most representative TFIIH views obtained upon averaging aligned images clustered into homogeneous classes. The stain-excluding protein densities are represented in white (upper panel) and are outlined by contours of equal density (lower panel). The six views correspond to different orientations of the molecule or to slight variations in staining. Each class average is identified by a number and the relative importance of the classes are indicated in percentages. The bar represents 100 nm in (A) and 20 nm in (B).
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4. Figure 3 Three-Dimensional Model of TFIIH Complex
(A) Diagram showing the orientations of the 100 class averages used in the final reconstruction. Each class-average is represented by a point in a (θ, φ) coordinate system. (B) Diagram representing the Fourier shell correlation function between two independent reconstructions (continuous line) and the 3σ threshold curve (dotted line). The vertical axis indicates the value of the Fourier shell correlation coefficient versus the resolution in 1/nm represented on the horizontal axis. (C) Surface representation of the 3D reconstruction of the human TFIIH at a resolution of 3.8 nm. This model includes the untilted images grouped into classes 1 to 5 shown in Figure 2B and their corresponding 60°-tilted views, which correspond to 94% of the data set. The model is turned around a vertical axis by increments of 40°. The bar represents 20 nm.
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5. Figure 4 Electron Microscopy Observation and Image Analysis of rIIH5
(A) Electron micrograph of rIIH5 molecules adsorbed on a carbon film and negatively stained with uranyl acetate. (B) Gallery of the most representative rIIH5 views. The stain-excluding protein densities are represented in white. The six views correspond presumably to different conformers of the complex since a continuum of structures is observed ranging from an almost linear structure (top left) to compact ring-like structures (bottom right). (C) Surface representation of a 3D model of rIIH5 calculated from the most homogeneous ring-shaped class (bottom right in Figure 4B) by a conical tilt series. The image population corresponds to about 10% of the data set of tilted and untilted molecular views. The model is turned around a vertical axis by increments of 30°. (D) Superimposition of the rIIH5 model onto that of TFIIH. The bar represents 140 nm in (A), 20 nm in (B), 13.5 nm in (C), and 3.4 nm in (D).
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6. Figure 5
Immunolabeling of TFIIH by Subunit-Specific Monoclonal Antibodies
In each row, panel 1 represents the closest reprojection of the 3D model of TFIIH, panel 2 shows the class average of unlabeled molecules whereas panel 3 represents the class average of labeled molecules. Panel 4 represents the difference map between the labeled and the unlabeled class averages. The antibody binding site is highlighted by an arrowhead in panel 5 on the surface representation of the 3D model of TFIIH. (A and B) Analysis of two independent TFIIH views labeled by Ab-cdk7. (C and D) Analysis of two independent TFIIH views labeled by Ab-XPD, which recognizes the C-terminal part of the protein. (E and F) Analysis of two independent TFIIH labeled by Ab-XPB, which recognizes the C-terminal part of the subunit. (G and H) Analysis of the binding of Ab-p44 to TFIIH. A single TFIIH view was obtained and two slightly different antibody binding sites are shown in the upper and lower rows.
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7. Figure 6 Model of the Quaternary Organization of TFIIH
The positions of subunits cdk7, XPD, XPB, and p44, as inferred from the immunolabeling experiments shown in Figure 5, are indicated by arrows on the three-dimensional model of human TFIIH. Note that the anti-XPB and -XPD antibodies are directed against the extreme C terminus of the proteins. The cyclin H, MAT1, and p34 subunits are tentatively positioned on the model according to well established subunit–subunit interaction studies. Additional data is required to position subunits p62 and p52.
Cell  , DOI: ( /S (00) )