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. 2017 Jan 6;6:e20070. doi: 10.7554/eLife.20070

Figure 1. Structural overview of the THO*•Sub2 complex.

(A) Schematic representation of THO and Sub2. Recombinant protein fragments that were used for crystallization are indicated by bars. The THO core complex (THO*) contains S. cerevisiae Tho2, Hpr1, Mft1, and Thp2, as well as S. bayanus Tex1 (81% sequence identity with S. cerevisiae Tex1). The DEAD-box ATPase Sub2 contains two recombinase A (RecA)-like domains labeled as Sub2-N and Sub2-C. (B) Coomassie-stained SDS-polyacrylamide gel electrophoresis (PAGE) of purified THO* complex and full-length Sub2. (C) Three views of the THO*•Sub2 complex with THO* in a cartoon representation and Sub2 in a surface representation. (D) Experimental electron density map of Sub2 after density modification, contoured at 1.0σ level corresponding to the view of the upper left panel in C.

DOI: http://dx.doi.org/10.7554/eLife.20070.002

Figure 1.

Figure 1—figure supplement 1. Crystallization of the THO*•Sub2 complex.

Figure 1—figure supplement 1.

(A) Silver-stained SDS-PAGE analysis of washed crystals of the THO*•Sub2 complex, indicating the presence of stoichiometric amounts of each subunit. (B) Omit 2Fo-Fc electron density map of the THO*•Sub2 complex calculated by rigid body refinement of the model omitting two helices of THO*. Only density in the vicinity of the omitted helices is shown. The map is contoured at 1.0σ. The region of the omit map is indicated by the box on the right side overall view.
DOI: http://dx.doi.org/10.7554/eLife.20070.003
Figure 1—figure supplement 2. Subunit architecture of THO.

Figure 1—figure supplement 2.

(A) Negative stain EM localization of THO subunits using N-terminal or C-terminal MBP tags. Four representative class averages are shown for each MBP tagged complex; one class average of the untagged THO* in the same orientation is shown as a control. The tag position is indicated by an arrowhead in each class average. Each box is approximately 44 × 44 nm. (B) Subunit localization indicated in the low-pass filtered THO* structure at 40 Å (Tang et al., 2007), according to A.
DOI: http://dx.doi.org/10.7554/eLife.20070.004