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. Author manuscript; available in PMC: 2019 Nov 1.
Published in final edited form as: Am J Med Genet A. 2018 Sep 14;176(11):2404–2418. doi: 10.1002/ajmg.a.40482

Fig. 5: WBS-linked Arg-378-Pro variant in DDX11.

Fig. 5:

(A) Location of disease-linked DDX11-R378P variant with respect to conserved helicase core motifs and Fe-S cluster. Conserved helicase motifs are shown in red and Fe-S motif is highlighted in yellow. Previously published variants in DDX11 shown in black and the novel variants from this series shown in blue. (B) Sequence alignment (Muscle Multiple Sequence Alignment) of region spanning R378 residue of DDX11 and related Fe-S containing DNA helicases XPD, FANCJ, and RTEL-1. (C) Coomassie-stained SDS-PAGE gel to analyze FLAG-M2 resin affinity-purified FLAG-tagged DDX11-WT and DDX11-R378P recombinant proteins expressed in 293T cells. E-elution. The intact full-length DDX11-R378P protein was hardly detectable in the eluted fraction compared to the full-length DDX11-WT protein. (D) Western blot analysis and quantification of DDX11 protein samples. Panel A. Sup- the supernatant obtained after centrifugation of cell lysates; Elution-Proteins obtained after 3X FLAG peptide elution in the final stage of purification. The quantification shows ~5-fold greater in wild-type DDX11 protein compared to the R378P DDX11 protein. (E) Western blot analysis of whole cell lysate protein from HeLa cells expressing recombinant DDX11-WT or DDX11-R378P. Actin serves as a loading control.